HK1213991B - Specimen processing systems and methods for preparing reagents - Google Patents

Description

Specimen processing system and method for preparing reagents

Cross Reference to Related Applications

This application claims benefit and priority from U.S. provisional patent application No. 61/746,085 filed on 26/12/2012 and U.S. provisional patent application No. 61/799,098 filed on 15/3/2013, both of which are incorporated herein by reference in their entirety.

Technical Field

The present disclosure relates to a system for preparing a specimen for analysis. In particular, the present disclosure relates to specimen processing systems and methods of processing specimens.

Background

A wide variety of techniques have been developed for preparing and analyzing biological specimens. Exemplary techniques include: microscopy, microarray analysis (e.g., protein and nucleic acid microarray analysis), and mass spectrometry. The sample for analysis is prepared by applying one or more liquids to the sample. If a specimen is treated with multiple liquids, the application and subsequent removal of the various liquids is important to produce a sample suitable for analysis.

Microscope slides bearing biological specimens (e.g., tissue sections or cells) are typically treated with one or more dyes or reagents to increase the color and contrast of the transparent or invisible cells or cellular components. Specimens for analysis can be prepared by manually applying dyes or other reagents to slides bearing the specimens. The labor intensive process often results in inconsistent processing due to individual techniques among laboratory technicians.

"dipping" automated machinery dips specimens into a liquid by a technique similar to the manual dipping technique. These automated machines are capable of processing specimens in batches by immersing racks carrying microscope slides into open baths. Unfortunately, liquid residue between the containers leads to contamination and degradation of the process liquid. Worse still, the detachment of cells from the slide carrying the specimen may contaminate other slides in the cuvette. These types of processes also utilize excessively large volumes of liquid, resulting in relatively high processing costs when reagents must be changed to reduce the likelihood of sample cross-contamination. Open containers are also susceptible to evaporative loss and oxidative degradation of reagents that can significantly alter reagent concentration and effectiveness, resulting in inconsistent processing. Processing of samples can be difficult without generating a significant volume of waste material that may require special handling and disposal.

Immunohistochemistry and in situ hybridization staining procedures are often used to prepare tissue specimens. The rate of immunohistochemical and in situ hybridization staining of sectioned fixed tissue on a microscope slide is limited by the rate at which molecules (e.g., conjugated biomolecules) can diffuse into the fixed tissue from aqueous solutions in direct contact with the tissue section. Tissue is often "fixed" immediately after resection by placing it in a 10% formalin solution that protects the tissue from autocatalytic destruction by cross-linking with most proteins via methylene bridges. The cross-linked tissue may present a number of additional diffusion barriers, including a bilayer lipid membrane that coats individual cells and organelles. The conjugated biomolecules (antibody or DNA probe molecules) can be relatively large, from a few kilodaltons to several hundred kilodaltons, which constrains them to diffuse slowly into solid tissues, with typical events required for adequate diffusion being minutes to hours. Typical incubation conditions are 30 minutes at 37 degrees celsius. The rate of staining is often driven by a concentration gradient and can therefore be increased by increasing the concentration of conjugate in the reagent to compensate for slow diffusion. Unfortunately, conjugates tend to be very expensive, and therefore, increasing their concentration is very wasteful and often economically not feasible. Furthermore, when high concentrations are used, excess conjugate driven into the tissue is retained in the tissue, is difficult to wash away, and results in high levels of non-specific background staining. To reduce noise due to non-specific background staining and increase the signal for specific staining, low conjugate concentrations and long incubation times are often used to allow the conjugate to bind only to specific sites.

Histological staining instruments often use relatively large volumes of reagent (100 μ L) in a pool of buffer, typically 300 μ L. Some conventional instruments mix the reagents by staggering tangential air jets onto an overlying oil layer that rotates and counter-clockwise when contacted by the staggered air jets, thereby imparting motion to an underlying aqueous pool. The mixing is slow and not particularly vigorous and can produce substantial evaporative losses, especially at elevated temperatures, which are often necessary. A large volume of wash liquid is used to physically displace a large pool of reagent covered with oil. This flushing procedure produces large volumes of waste liquid, which can be hazardous waste.

Disclosure of Invention

Some embodiments of the present technology relate to an automated slide processing apparatus for dispensing a liquid onto one or more microscope slides. In one embodiment, the automated slide processing apparatus can comprise: a carousel comprising a plurality of storage wells; and a reagent pipette assembly comprising a reagent pipette movable between at least one loading position for obtaining reagent from one of the reservoir wells and at least one dispensing position for dispensing reagent onto one of the microscope slides. In some arrangements, the automated slide processing apparatus can further comprise: a wash pipette assembly configured to wash the plurality of reservoir wells; and a drive mechanism coupled to the carousel and configured to rotate the carousel to position the reservoir wells relative to the reagent pipette assembly and/or the wash pipette assembly.

At least some of the embodiments of the automated slide processing apparatus can include: a filling station comprising a plurality of containers containing reagents; and a plurality of slide treatment stations. For example, the reagent pipette assembly can be movable through a lumen of the automated slide processing apparatus to transport reagents obtained at a filling station to a carousel and dispense reagent mixtures from the carousel onto one of the microscope slides. In another embodiment, the reagent pipette assembly is movable between a filling position for obtaining reagent from a container at a filling station and a dispensing position for filling one or more of the storage wells with reagent from the filling station. In some embodiments, the automated slide processing apparatus has a hybrid mode in which a reagent pipette assembly mixes reagents within one or more of the reservoir wells and dispenses the reagent mixtures onto a microscope slide. In other embodiments, the wash pipette assembly mixes the reagents within one of the reservoir wells.

For example, the drive mechanism can be configured to sequentially rotate the reservoir wells beneath the wash pipette of the wash pipette assembly and/or the reagent pipette of the reagent pipette assembly. In one embodiment, the reagent pipette assembly has a reagent loading state for obtaining reagent from the reservoir wells while the wash pipette assembly delivers, for example, wash solution to another of the reagent wells. In some embodiments, the wash pipette assembly includes a pipette movable into each of the reservoir wells. In another embodiment, the wash pipette assembly is fluidly coupled to a vacuum source and draws liquid from one of the reservoir wells when the vacuum source is drawing a vacuum. In some embodiments, the reagent pipette assembly accesses the reservoir well at the same location, and the carousel is capable of rotating the reservoir well to a location accessible to the reagent pipette assembly. In other embodiments, the carousel rotates to position the reagent wells such that the reagent pipette assembly approaches the reservoir wells at different locations.

In some embodiments, the carousel has a dedicated waste channel to direct liquid into the drain pipe without risk of contaminating other adjacent wells. In at least some embodiments of the present technology, the carousel includes spillways configured to allow fluids (e.g., cleaning fluids, reagents, etc.) to flow from the storage wells to prevent cross-contamination (e.g., the flow of fluids between adjacent storage wells). The spillways can have the same radial length to inhibit or prevent recirculation of waste streams into adjacent wells. In one embodiment, the carousel can include a plurality of overflow spacers each positioned circumferentially between adjacent storage wells. In one example, the spill barrier extends upwardly and radially inwardly from the storage well. In further embodiments, the carousel can include a drain and a spillway that allows overflow of the reagent to flow from the storage well toward the drain.

In one embodiment, the automated slide processing apparatus includes a controller communicatively coupled to the drive mechanism and configured to command the drive mechanism such that the drive mechanism sequentially moves each of the reservoir wells to a wash position for washing by the wash pipette assembly. In some embodiments, the controller stores and executes instructions for commanding the reagent pipette to sequentially fill the storage wells with reagent from the reagent containers. In another embodiment, an automated slide processing apparatus includes a controller having mixing instructions executable to command a reagent pipette assembly to deliver at least two reagents to one or more of the reservoir wells to produce a reagent mixture. In one arrangement of such an embodiment, the controller has mixed reagent dispensing instructions that are executable to command the reagent pipette assembly to dispense the reagent mixture onto the specimen.

Additional embodiments of the present technology relate to methods of sequentially delivering reagents to a plurality of storage wells of a carousel to produce a reagent mixture. The carousel can be rotatable to sequentially position the storage wells at one or more washing locations. The method can further include: the reagent pipette is at least partially filled with a reagent mixture from one of the storage wells when at least one of the storage wells is at the wash location (or locations). The reagent pipette assembly is capable of aspirating multiple reagents, in part, from one or more of the reservoir wells (premix) for single or multiple dispensing onto one or more slides. After at least partially filling the reagent pipette with reagent, the method can further include robotically moving the reagent pipette toward the microscope slide and dispensing reagent onto the microscope slide. In still other embodiments, the method can include rotating the carousel such that one of the storage wells containing reagent (e.g., excess or residual reagent) is located at a wash position, and washing the storage well at the wash position to remove the reagent.

At least some embodiments of the present technology relate to automated specimen processing systems capable of processing specimens carried on slides. At least some embodiments include an automated specimen processing system that includes a slide ejector assembly. The slide ejector assembly can include a slide staging device (stationary device) configured to receive a slide. The slide ejector assembly can further comprise: a slide alignment device configured to engage the slide at a plurality of contact points to move the slide from an unaligned position to an aligned position. In one embodiment, the slide alignment device can include: a first alignment member and a second alignment member located opposite the first alignment member. The first and second alignment members are movable between an open position for receiving a slide and a closed position for aligning and/or holding the slide.

In some embodiments, the first alignment member can include a first contact region and a second contact region for engaging a first edge of the slide. In some embodiments, the second alignment member can include: a third contact region for engaging a second edge of the slide opposite the first edge. In various embodiments, the slide alignment device is configured to engage the slide at three contact points. In one example, the point of contact can be a small discrete area of the slide that is contacted by one of the first, second, or third contact regions. In one embodiment, the slide can be moved from the unaligned position to the aligned position on the standby platform by pivoting the slide about a point (e.g., a middle point) between the three contact points. In another embodiment, moving the slide from the unaligned position to the aligned position includes: the slide longitudinal axis is aligned with the standby platform longitudinal axis.

In some embodiments, an over travel inhibitor and a slide holding region between the over travel inhibitor and the slide ejector are provided. The over travel inhibitor can be positioned to inhibit movement of the slide through the slide holding region, for example. In one embodiment, the over travel inhibitor includes a vacuum port positioned to draw a vacuum between the slide and the standby platform as the slide moves through at least a portion of the standby platform. In another embodiment, the over travel inhibitor can include: a sensor for detecting the presence of a slide on the standby platform.

At least some embodiments of an automated specimen processing system include: at least one specimen processing station and a transfer head configured to transport slides from the standby platform to the specimen processing station. In one embodiment, the transfer head can include a head alignment feature receivable by at least one of a corresponding alignment feature of the slide staging device and/or an alignment feature of the specimen processing station. In one embodiment, the head alignment feature includes a first alignment pin and a second alignment pin, and the corresponding alignment feature of the slide staging device includes a first opening and a second opening positioned to receive the first alignment pin and the second alignment pin, respectively. In further embodiments, the transfer head can comprise: a capture feature configured to engage the slide and transport the slide in an aligned position. For example, the capture features can include: a vacuum port positioned to draw a vacuum between the upper surface of the slide and the transfer head while the slide is being transported.

At least some embodiments of the automated specimen processing system include a controller communicatively coupled to the slide ejector assembly. The controller, for example, can be programmed to command the slide alignment device to move the first alignment feature toward the standby platform in a first direction and to move the second alignment feature toward the standby platform in a second direction opposite the first direction to engage the slide at the plurality of contact points to move the slide. The controller can be programmed to command the slide alignment device to move the first alignment device in the second direction and the second alignment member in the first direction to release the slide in the aligned position. In another embodiment, the controller can be programmed to control the transfer head to align with the slide staging device and transport the slide from the standby platform to the specimen processing station.

At least some embodiments of the present technology relate to an automated specimen processing system including a slide staging device and a transfer head. In one embodiment, the slide staging device can include a staging platform configured to receive a microscope slide and an alignment device having a first alignment member and a second alignment member positioned opposite the first alignment member. In some embodiments, the alignment device is configured to engage the microscope slide at a plurality of contact points, thereby moving the slide from the unaligned position to the aligned position. In some arrangements, the transfer head can be configured to transport microscope slides from the standby platform to the specimen processing station. For example, the transfer head can have a head alignment feature receivable by at least one of a corresponding alignment feature of the slide staging device and/or an alignment feature of the specimen processing station. In various embodiments, the first alignment member can have a first contact region and a second contact region for engaging a first edge of the slide, and the second alignment member can have a third contact region for engaging a second edge of the slide opposite the first edge.

Some embodiments of the present technology relate to methods of transporting specimen-bearing microscope slides in automated processing systems. In one embodiment, the method includes sequentially moving a plurality of specimen-bearing microscope slides from a carrier to a slide staging device. By engaging individual specimen-bearing microscope slides at multiple contact points, the individual specimen-bearing microscope slides can be aligned with the longitudinal axis at the slide staging device. Optionally, after moving individual specimen-bearing microscope slides from the carrier to the slide staging device, a vacuum is drawn through the over-travel inhibitor to capture the specimen-bearing microscope slides on a standby platform of the slide staging device and detect the presence of the slides on the standby platform. In some embodiments, the method further includes transporting individual specimen-bearing microscope slides from the slide staging device to one or more specimen processing stations.

In some embodiments, transporting individual specimen-bearing microscope slides includes: the transfer head of the transport assembly is aligned with the slide staging device and individual specimen-bearing microscope slides are picked up from the slide staging device while maintaining the aligned position. In other embodiments, the alignment features of the transport assembly can be aligned with corresponding alignment features at the slide staging device prior to transporting individual specimen-bearing microscope slides. In further embodiments, transporting the individual specimen-bearing microscope slides includes drawing a vacuum between the individual specimen-bearing slides and a transport assembly, and the transport assembly is configured to transport the specimen-bearing slides to one or more specimen processing stations.

At least some embodiments of the present technology relate to an automated slide processing apparatus configured to apply at least one reagent to a specimen carried by a microscope slide. The slide treatment station can include: a support element having a support surface, at least one port, and a sealing member having a non-circular shape (e.g., when viewed from above). The sealing member is movable between an uncompressed state and a compressed state. In the uncompressed state, the sealing member can extend upwardly beyond the bearing surface. In the compressed state, the sealing member can be configured to: maintaining a seal with a back side of the microscope slide while the microscope slide is resting against the support surface by a vacuum drawn through the at least one port. In some embodiments, the sealing member can have a rounded rectangular shape (e.g., a shape having rounded corners with a radius less than the length of a straight side) or a rectangular shape when viewed from above. In one embodiment, the sealing member has a rounded polygonal shape or a polygonal shape when viewed along an axis substantially perpendicular to the support surface.

In some embodiments, at least a portion of the support element can have a non-circular shape and can extend between the sealing member and the at least one vacuum port. In one embodiment, the support element comprises a groove and the sealing member comprises a flexible gasket having a body and a lip. The body can be located in the groove, and the lip can extend radially outward from the body. In some embodiments, the lip is movable between a compressed configuration and an uncompressed configuration. In the uncompressed configuration, the lip can extend upwardly from the channel. In the uncompressed configuration, the lip can extend toward the side wall of the groove. In one embodiment, the lip is movable between an uncompressed configuration and a compressed configuration without contacting the sidewalls of the groove. The lip can be spaced from, but can physically contact, a sidewall of the groove when the microscope slide is placed against the support surface to inhibit movement of the microscope slide relative to the support element. In one embodiment, the lip is sufficiently stiff to prevent any rotation of the slide about the vertical axis. In this way, the slide is rotationally fixed relative to the support surface. In one embodiment, the lip is configured to physically contact the sidewall when the microscope slide is rotated at least about 2 degrees about the vertical axis.

The sealing member in the compressed configuration can be located on one side of a plane of the backside surface of the microscope slide when the microscope slide is placed against the support surface. In the uncompressed configuration, the sealing members can be located on both sides of the plane. The support element can include a vacuum surface surrounded by at least one vacuum port. The vacuum surface can be spaced from and located below the plane such that the vacuum surface and the microscope slide at least partially define a vacuum chamber having a height that is less than a height of the sealing member.

In some embodiments, the sealing member can include a lip configured to deflect primarily in a direction perpendicular to the backside surface of the microscope slide during use. The lip is movable between an uncompressed configuration for contacting a slide moving toward the support surface and a compressed configuration for maintaining the hermetic seal. In an uncompressed state, the lip can extend upwardly beyond the bearing surface. In the compressed state, the lip can be located at or below the bearing surface. In some embodiments, the lip can be configured to deflect as the microscope slide moves toward the support surface to form an airtight seal with the slide. In some embodiments, the sealing member can be positioned to underlie a label of a microscope slide during use.

At least some embodiments of the automated slide processing systems include a vacuum source in communication with the at least one vacuum inlet and configured to draw sufficient vacuum to maintain the hermetic seal. In some embodiments, the slide processing system can include a heater configured to heat the support element, thereby causing the support element to conductively heat the microscope slide while the sealing member maintains the hermetic seal.

Some embodiments of the present technology relate to methods of holding a microscope slide. In one embodiment, the method comprises: positioning a microscope slide on a support element, the support element having a first portion surrounding a second portion; and aligning the label-bearing portion of the slide with the second portion and the specimen-bearing portion of the slide with the first portion. The method further comprises the following steps: a vacuum is drawn through the second portion and the slide is sealed to the sealing member. In some embodiments, the method further comprises: inhibiting at least one of translational or rotational movement of the slide relative to the support element. The method further comprises the following steps: a vacuum chamber is created between the second portion and the back side of the microscope slide. In some embodiments, positioning the microscope slide comprises: contacts a top of the sealing member and deflects the top of the sealing member in a direction substantially perpendicular to a path of travel of the slide.

At least some embodiments of the present technology relate to biological specimen processing systems capable of processing specimens carried on slides. The specimen processing system is capable of sequentially delivering slides and opposable (opposable) to a specimen processing station. The specimen processing station can use the opposable to manipulate and direct a series of liquids to the specimen. The liquid can be manipulated on and/or across the surface of the slide in conjunction with capillary action while the specimen processing station controls the processing temperature of histological staining, immunohistochemical staining, in situ hybridization staining, or other specimen processing protocols. In some embodiments, opposable is a surface or opposable element(s) capable of manipulating one or more substances on a slide. Manipulating a substance in a watch fluid form can include: diffusing the fluid; displacing a film of fluid, or changing a dose, band or film of fluid.

At least some embodiments of the present technology relate to a system for contacting a biological specimen with a liquid by moving an opposable in contact with the liquid. The non-planar (e.g., curved) wetted surface of the opposable is separated from the specimen-bearing slide by a distance sufficient to form a meniscus layer of liquid between the wetted surface and the slide. The meniscus layer contacts at least a portion of the biological specimen and moves across the slide using capillary action and other manipulation actions.

In some embodiments, the meniscus layer can be a relatively thin fluid film, fluid ribbon, or the like. The opposable can be moved to different positions relative to the slide and can accommodate different volumes of liquid forming the meniscus layer. Capillary action can include, but is not limited to: movement of the meniscus layer due to the phenomenon of adhesion, cohesion and/or surface tension liquids spontaneously passing through the gap between the curved wet opposable surface and the slide. The opposable can manipulate (e.g., agitate, displace, etc.) the liquid to process the specimen using a relatively small volume of liquid to help manage waste and provide consistent processing. Evaporation losses (if any) can be managed to maintain desired liquid volumes, reagent concentrations, etc. The specimen can be processed using a relatively small volume of liquid in order to reduce liquid waste.

In some embodiments, the system includes one or more automated slide holders that can heat individual slides via conduction to generate a temperature profile across the slides that compensates for heat loss. Heat loss can result from evaporation of liquid in a gap between the slide and an opposable disposed proximate to the slide. In one embodiment, the slide holder has a slide-supporting surface and produces a non-uniform temperature profile along the slide-supporting surface that contacts the slide such that the specimen-bearing surface of the slide has a substantially uniform temperature profile when the slide is positioned on the slide-supporting surface. In some embodiments, a non-uniform temperature profile is generated across the slide support surface while a substantially uniform temperature profile is generated along the mounting surface of the slide. Another feature of at least some embodiments of the technology is: the slide holder can be configured to create a low temperature heating zone and a high temperature heating zone surrounding the low temperature heating zone. The high temperature zone can compensate for higher evaporative heat losses to maintain the specimen at a substantially uniform temperature.

At least some embodiments include a specimen processing system including a slide ejector assembly for removing slides from a slide carrier. The slide ejector assembly includes a carrier, a slide staging device, and an actuator assembly. The carrier handler is configured to receive and hold a slide carrier that holds a plurality of slides. The slide staging device includes a standby platform and a slide alignment device configured to move a slide from an unaligned position to an aligned position at the standby platform. The actuator assembly includes: a slide ejector positioned to move relative to the slide carrier to transfer individual slides from the slide carrier to the standby platform. The slide can thus be transferred to the standby platform without the use of, for example, a mechanical gripper or suction cup device that pulls the slide from one location to another.

In some embodiments, the carrier handler is configured to move the slide carrier relative to the slide ejector to sequentially stage one of the slides for delivery to the standby platform. In some embodiments, the carrier handler includes a carrier receiver and a receiver rotator. The receiver rotator is capable of rotating the slide carrier from a vertical slide orientation to a horizontal slide orientation. In one embodiment, the carrier handler comprises: a carrier receiver movable between a loading position for loading a slide carrier and a slide unloading position. The carrier handler can include a receiver rotator and a transport device. A receiver rotator is coupled to the carrier receiver and is operable to move a slide carrier held by the carrier receiver from a vertical slide orientation to a horizontal slide orientation. The transport device is configured to move the slide carrier oriented along the horizontal slide vertically between the slide ejector and the standby platform.

In some embodiments, the slide staging device includes an ejector stop positioned to prevent the slide ejector from moving past the end of the slide holding area of the standby platform. The slide ejector is movable from a first position to a second position. In some embodiments, the slide ejector moves through the slide carrier to push a slide out of the slide carrier.

The standby platform can include a slide holding region and an over travel inhibitor. The slide holding region is positioned between the over travel inhibitor and the slide ejector. The slide ejector is positioned to move the slides one at a time from the slide carrier toward the over travel inhibitor. In some embodiments, the over travel inhibitor includes a vacuum port positioned to draw a vacuum between the slide and the standby platform as the slide is moved through at least a portion of the standby platform by the slide ejector.

In some embodiments, the slide alignment device includes a pair of jaws movable between an open position for receiving a slide and a closed position for aligning the slide. In one embodiment, the gripper centers the slide relative to the raised slide holding area of the standby platform when the gripper is moved from the open position to the closed position.

The actuator assembly includes a reciprocating drive mechanism coupled to the slide ejector and configured to move the slide ejector so as to eject a slide from the slide carrier and onto the standby platform. In some embodiments, the slide ejector is movable through a slide carrier receiving gap between the actuator assembly and the slide staging device.

In some embodiments, the specimen processing system can further include one or more specimen processing stations and one or more transfer heads. The transfer head can be configured to transport the slide from the standby platform to one of the specimen processing stations. In some embodiments, the at least one transfer head can have a head alignment feature receivable by at least one of an alignment feature of the slide staging device and/or an alignment feature of the specimen processing station. In some embodiments, the head alignment feature includes a first alignment pin and a second alignment pin. The alignment feature of the slide staging device can include a first opening and a second opening. The first and second openings are positioned to receive the first and second alignment pins, respectively. In some embodiments, the alignment feature of the specimen processing station can include first and second openings, and the first and second openings are positioned to receive first and second alignment pins, respectively, of the head alignment feature.

In some embodiments, the specimen processing system can further include a controller communicatively coupled to the slide ejector grouping. The controller can be programmed to command the actuator assembly to move a first slide located below a second slide from the slide carrier to the standby platform and to move the second slide to the standby platform after moving the first slide to the standby platform.

In some embodiments, a method of transporting a specimen-bearing microscope slide comprises: a carrier containing a plurality of specimen-bearing microscope slides is delivered to an ejector assembly. The carrier is moved toward a slide staging device of the ejector assembly. The specimen-bearing microscope slides are sequentially moved from the carrier to the slide staging device. The slide staging device moves from the receiving slide configuration to the aligning slide configuration to move individual specimen-bearing microscope slides to an aligned position at the slide staging device. Individual specimen-bearing microscope slides are transported from the slide staging device of the ejector assembly to one or more specimen processing stations.

In some embodiments, the carrier is rotatable to move the plurality of specimen-bearing microscope slides from the first orientation to the second orientation. In some embodiments, the first orientation is a substantially vertical orientation and the second orientation is a substantially horizontal orientation.

In some embodiments, a specimen-bearing microscope slide can be sequentially moved from the carrier to the slide staging device by pushing the specimen-bearing microscope slide onto and along the slide staging device. Additionally or alternatively, the lowermost specimen-bearing microscope slide is held by a carrier to the slide staging device. The process can be repeated until most or all of the slides have been removed from the slide carrier.

In certain embodiments, individual specimen-bearing microscope slides can be carried from the slide staging device to a specimen processing station configured to individually process specimen-bearing microscope slides. Additionally or alternatively, the specimen-bearing microscope slides can be sequentially moved from the carrier to the slide staging device by moving a first specimen-bearing microscope slide from the carrier to the slide staging device. After the first specimen-bearing microscope slide is transported away from the slide staging device, a second specimen-bearing microscope slide is transported from the carrier to the slide staging device.

In some embodiments, the slide staging device can be moved from the receive slide configuration to the align slide configuration by moving the pair of jaws from the open position to the closed position to contact and move a specimen-bearing microscope slide located between the jaws from the unaligned position to the aligned position. In certain embodiments, the gripper can center the slide relative to a raised portion of the slide staging device on which the slide rests.

In some embodiments, specimen-bearing microscope slides are sequentially removed from the carrier by (a) pushing the specimen-bearing microscope slide at the slide ejection position such that the specimen-bearing microscope slide moves onto the slide staging device, and (b) repeating process (a) until the carrier is empty. In one embodiment, an elongated ejector moves through a carrier (e.g., a basket) to push slides onto a slide staging device.

A vacuum can be drawn between individual specimen-bearing microscope slides and the slide staging device. For example, a sufficient vacuum can be drawn to inhibit or limit movement of the slide along the slide staging device. The vacuum can be reduced or eliminated to remove the slide from the slide staging device.

In some embodiments, the carrier is a slide rack that includes shelves that hold specimen-bearing microscope slides in a spaced-apart arrangement. The specimen-bearing microscope slides can be sequentially moved from the carrier to the slide staging device by indexing the rack at a slide removal position adjacent to the platform of the slide staging device. In some embodiments, the slide at the slide removal position is slightly higher than the slide staging device.

Specimen-bearing microscope slides can be sequentially removed from the carrier by (a) reciprocating a slide ejector between an initial position and an ejected position to move at least one of the specimen-bearing microscope slides from the carrier to the slide staging device, and (b) repeating process (a) to remove at least a majority of the specimen-bearing microscope slides from the carrier. In some embodiments, a slide ejector is used to remove all specimen-bearing microscope slides from the carrier.

In some embodiments, a slide processing apparatus for processing a specimen carried by a slide includes a staining module. The staining module includes a slide holder platen, an opposable element, and an opposable actuator. The slide holder platen has a first sidewall, a second sidewall, and a slide receiving area between the first sidewall and the second sidewall. The slide is positioned on the slide receiving area. The carrier sheet includes a first edge and an opposing second edge. The opposable element is disposed proximate the slide and includes a first edge portion and an opposing second edge portion. The opposable actuator holds the opposable element to form a capillary gap between the opposable element and the slide. The first edge portion of the opposable element is closer to the first sidewall than the first edge of the slide. The second edge portion of the opposable element is closer to the second sidewall than the second edge of the slide.

In some embodiments, the slide processing apparatus includes a dispenser positioned to deliver a supplemental liquid therebetween while holding the liquid in a gap between the opposable element and the slide. Further, the slide processing apparatus can include a controller communicatively coupled to the dispenser and programmed to command the dispenser to deliver the supplemental liquid to maintain a volume of liquid between the opposable element and the slide within the equilibrium volume range. In some embodiments, the controller is programmed to deliver the supplemental liquid at a predetermined rate. In one embodiment, the predetermined rate is equal to or less than about 110 μ L per minute at a temperature of about 37 ℃ for the bulk liquid. In one embodiment, the predetermined rate is equal to or less than about 7 μ L per minute at a temperature of about 37 ℃ for non-bulk liquids. The rate can be selected based on the specimen staining protocol being processed.

In some embodiments, the slide treatment apparatus further includes a plurality of additional staining modules and a controller configured to independently control each staining module. The staining module can use disposable or reusable opposable elements to spread and move reagents through the specimen.

The first edge portion of the opposable element can extend past the first edge of the slide toward the first sidewall. A second edge portion of the opposable element can extend past a second edge of the slide toward the second sidewall. The opposable element can include a mounting end having at least one slot sized to be received and retained by at least a portion of the opposable actuator. In some embodiments, the opposable element has a suction end and an arcuate body extending from the suction end. The arcuate body is configured to roll along the slide to move the liquid across the surface of the slide. The suction end has a radius of curvature equal to or less than 0.08 inches. Other sizes can also be used.

The staining module can include: at least one heating element positioned to conductively heat the first sidewall, the second sidewall, or both. The opposable actuator is movable to roll the curved portion of the opposable element along the slide to move the band of liquid across at least a portion of the slide carrying the specimen. The first and second sidewalls can be used to heat the slide, specimen, and/or liquid as the band of liquid is manipulated through the specimen.

In some embodiments, the slide processing apparatus can include a contact surface of the slide receiving region that supports the slide such that an edge portion of the slide extends outward from an edge of the opposable.

In some embodiments, a system for processing a specimen carried by a slide includes a specimen processing station and a controller. The specimen processing station includes an opposable actuator and a slide holder platen. The slide holder platen includes a slide support region and a liquid replenishment device. The slide holder platen is configured to heat liquid on the slide at the slide support region when the opposable element held by the opposable actuator contacts and moves the liquid across the slide surface. The replenishment device is configured to deliver a replenishment liquid between the opposable element and the slide. The controller is programmed to control the specimen processing station such that the replenishment device delivers a replenishment liquid at a replenishment rate to compensate for evaporative loss of the liquid.

In some embodiments, the controller includes one or more memories and a programmable processor. The memory stores a first sequence of program instructions and a second sequence of program instructions. The programmable processor is configured to execute a first sequence of program instructions to treat a specimen on the slide with a first liquid and to execute a second sequence of program instructions to treat the specimen with a second liquid different from the first liquid. In some embodiments, the programmable processor is configured to execute a first sequence of program instructions to heat the slide to a first temperature using the slide holder platen, and the controller is configured to execute a second sequence of program instructions to heat the slide to a second temperature using the slide holder platen, the second temperature being different from the first temperature.

In some embodiments, the controller is configured to execute a first sequence of program instructions to command the supplemental device to deliver the first liquid to the slide at a first rate. The controller is further configured to execute a second sequence of program instructions to command the supplemental device to deliver a second liquid to the slide at a second rate different from the first rate. In a particular embodiment, the first rate corresponds to an evaporation rate of the first liquid and the second rate corresponds to an evaporation rate of the second liquid. The controller can help mitigate evaporative losses.

In some embodiments, the controller includes a memory that stores a replenishment program executable by the controller to maintain the volume of liquid on the slide within the equilibrium volume range. In particular embodiments, the equilibrium volume ranges from about 70 μ L to about 260 μ L. In a particular embodiment, the controller is programmed to command the specimen processing station to maintain the liquid volume between a maximum equilibrium volume corresponding to an over-wet condition and a minimum equilibrium volume corresponding to an under-wet condition. In some embodiments, the controller is programmed to command the specimen processing station to move a volume of liquid through the specimen held on the slide by moving an opposable element held by an opposable actuator relative to the slide, and is further programmed to deliver supplemental liquid from the replenishment device to substantially compensate for the reduction in volume of liquid due to evaporation.

In some embodiments, the controller is configured to receive reference evaporation rate information (e.g., evaporation rate information of the liquid) from the memory and to control the specimen processing station based on the reference evaporation rate information. Additionally or alternatively, the controller can be programmed to command the specimen processing station such that the replenishment device provides the replenishment liquid at a rate selected based on the evaporation rate of the liquid.

In some embodiments, the system for processing a specimen further comprises an opposable element and a controller. The opposable element is held by the opposable actuator and is configured to extend outwardly past an edge of the slide. The controller is programmed to: when the evaporation rate of the liquid remains equal to or less than about a predetermined rate (e.g., 7 μ L per minute, 5 μ L per minute, etc. at a temperature of about 37 ℃), the specimen processing station is controlled to move the opposable element while the opposable element manipulates the liquid across the slide.

In some embodiments, the slide holder platen includes a heating element that receives electrical energy and outputs thermal energy to heat the slide via conduction. The heating element can comprise one or more resistive heating elements.

In some embodiments, a method for processing a specimen carried by a slide includes: the liquid on the slide held by the slide holder is heated. The opposable element is rolled to contact the liquid on the slide and move the liquid through the biological specimen on the slide. The replenishment rate is determined based on the evaporation rate of the liquid. Delivering a makeup liquid based on the replenishment rate to substantially compensate for evaporative loss of liquid. The opposable element (which contacts the liquid including the compensation liquid) is rolled so as to repeatedly contact the specimen with the liquid.

The volume of the supplemental liquid delivered onto the slide can be equal to or greater than the volume of liquid reduced via evaporation. Additionally or alternatively, the supplemental liquid can be delivered onto the slide by delivering the supplemental liquid to maintain a volume of liquid on the slide equal to or greater than a minimum equilibrium volume or equal to or less than a maximum equilibrium volume. Additionally or alternatively, the supplemental liquid can be delivered onto the slide while the opposable element is rolling along the slide.

In some embodiments, a method of processing a specimen on a slide includes: the liquid is moved along the slide using opposable elements that contact the liquid. The temperature of the liquid on the slide is controlled while the liquid is moved. At least one of a volume of the liquid and/or a total evaporation rate of the liquid is evaluated, and a supplemental liquid is delivered onto the slide based on the evaluation to maintain the volume of the liquid on the slide within an equilibrium volume range. In particular embodiments, the volume of liquid and the total evaporation rate of the liquid may be received from a reservoir to evaluate the volume of liquid and the total evaporation rate of the liquid from the reservoir, the reservoir evaluating the volume of at least one liquid and/or the total evaporation rate of the liquid including receiving. The equilibrium volume range can be about 125 μ L to about 175 μ L.

In some embodiments, the slide processing apparatus includes a slide holder platen and an opposable actuator. The slide holder platen has a receiving area configured to receive a slide with a first side of the slide facing the receiving area and a second side facing away from the receiving area. The opposable actuator is positioned to hold the opposable element to define a capillary gap between the opposable element and a slide at the receiving region. The opposable actuator is configured to: moving the capillary gap along the slide in a first direction to move the band of liquid from a first position to a second position across the length and width of the second side of the slide; and narrowing the band of liquid (e.g., reducing the width of the band of liquid in a direction substantially parallel to the first direction).

In some embodiments, the opposable actuator is configured to alternately roll the opposable element along the slide in a first direction and a second direction opposite the first direction to manipulate the band of liquid across the surface of the slide between the first position and the second position. A band of liquid at a first location between one end of the opposable element and the slide; and the band of liquid at the second position is between the opposable element and an end of the slide. The band of liquid can be narrowed at each of the first and second locations before moving the band of liquid to the other of the first and second locations. In some embodiments, the opposable actuator is a variable bandwidth compression opposable actuator configured to reduce a width of the band of liquid by a predetermined amount. The predetermined amount can be selected by a controller or an operator.

In some embodiments, the opposable actuator is configured to move the opposable element relative to the slide to reduce a width of the band of liquid at an end of an opening defined by the slide and/or at least one of the opposable elements by 50%, 40%, or 25%. Additionally or alternatively, the opposable actuator can be configured to move the opposable element to displace the band of liquid between the first position and the second position while maintaining a latitudinal width of the band of liquid. In some embodiments, the opposable actuator is movable between a first configuration in which the band of liquid narrows at a first end of an opening between the opposable element and an end of the slide, and a second configuration in which the band of liquid narrows at a second end of the opening. In some embodiments, the opposable actuator can be moved to the over-rolling configuration to move the first side of the band of liquid toward the second side of the band of liquid to reduce the width of the band of liquid while substantially statically holding the second side of the band of liquid at an end of one of the opposable element and the slide.

In some embodiments, the slide processing apparatus further includes a staining module and a controller. The staining module includes a slide holder platen and an opposable actuator. The controller is communicatively coupled to the staining module. The controller is programmed to command the staining module to move the opposable element to move the capillary gap.

In some embodiments, the slide processing apparatus further includes an opposable element including a mounting end held by the opposable receiver of the opposable actuator, an adsorption end opposite the mounting end, and a body. The body is located between the mounting end and the adsorption end. The absorbing end cooperates with the slide as the mounting end is moved away from the slide to accumulate liquid at an end of the mounting surface of the slide proximate to the label on the slide.

In some embodiments, the slide processing apparatus further comprises: an opposable element having a tapered end facing the receiving area. The tapered end is positioned to contact and attract the band of liquid. In a particular embodiment, the tapered end comprises: a rounded region extending between the opposed longitudinally extending edges of the opposable element.

In some embodiments, the opposable actuator has a rolling state that rolls the opposable element along the slide to move the band of liquid from a position at one end of an opening defined by one end of the slide and the opposable element to a position at an opposite end of the opening. The opposable actuator can have a static state that holds the opposable element stationary relative to the slide to perform, for example, culturing.

In some embodiments, the slide processing apparatus further comprises: a slide supported by the contact surface of the receiving region such that the slide extends laterally outward past opposite edges of the contact surface. The slide can carry one or more specimens.

In some embodiments, the slide processing apparatus further comprises: an opposable element held by an opposable actuator. The opposable element has a curved attracting end. The suction end can have a radius of curvature equal to or less than about 0.08 inches. In a particular embodiment, the opposable element has an arcuate body for rolling along the slide at the receiving area.

In some embodiments, the slide processing apparatus includes a slide holder platen and an opposable actuator. The opposable actuator includes an opposable receiver and a drive mechanism. The opposable actuator is positioned to hold the opposable element to define a capillary gap between the opposable element and a slide held by the slide holder platen. The drive mechanism has a rolling state for rolling the opposable element in a first direction along the slide to move the liquid band to one end of the space between the opposable element and the slide. The drive mechanism has an over-rolling condition for rolling the opposable element in a first direction to reduce a width of the band of liquid attracted at the end of the space.

In some embodiments, the opposable actuator is configured to move the opposable element to move the band of liquid across at least a majority of the mounting surface of the slide. The width of the band of liquid can be reduced by moving at least a portion of the opposable element away from the slide. The width of the band of liquid is in a direction substantially parallel to the longitudinal axis of the slide.

In some embodiments, a method for processing a specimen carried by a slide includes: delivering the slide and the opposable element to a staining module. The opposable element held by the staining module is positioned relative to a slide held by the staining module to hold the liquid in a capillary gap between the slide and the opposable element. The opposable element is moved relative to the slide to displace the liquid in a first direction substantially parallel to a longitudinal axis of the slide toward an end of the opening between the slide and the opposable element. The opposable element moves relative to the slide to reduce a width of the band of liquid in a first direction when the band of liquid is attracted at the end of the opening.

In some embodiments, the band of liquid is moved alternately between one end of the opening and an opposite end of the opening by rolling the opposable element along the slide in a first direction and a second direction opposite the first direction. The opposable element can include: one or more gap elements for maintaining a spacing between the body of the opposable element and the slide.

In some embodiments, the band of liquid is diffused to increase the width of the band of liquid. The diffused band of liquid can be moved across the specimen on the slide. In a particular embodiment, the width of the liquid band is reduced at one end of the capillary gap, after which the liquid band is moved to the other end of the gap.

In some embodiments, the method for processing a specimen further comprises drawing substantially all of the liquid at the end of the gap while reducing the width of the band of liquid.

In some embodiments, the method for processing a specimen further comprises: the liquid band is displaced across the specimen on the slide while maintaining the width of the liquid band.

In some embodiments, the method for processing a specimen further comprises: the width of the band of liquid is reduced by at least 50% by moving the opposable element relative to the slide. The volume of liquid can be equal to or greater than about 75 μ L.

In some embodiments, the width of the band of liquid is less than the length of the band of liquid. The width of the band of liquid is substantially parallel to the longitudinal axis of the slide. The length of the band of liquid is substantially perpendicular to the longitudinal axis of the slide.

In some embodiments, the slide heating apparatus includes a support element and a heater. The support element has a support surface that supports a slide with a back surface facing the support surface and a specimen-bearing surface of the slide opposite the back surface of the slide. The heater is coupled to the support element. The slide heating apparatus is configured to deliver thermal energy non-uniformly through the support surface to the back surface of the slide via conduction to substantially compensate for non-uniform heat loss associated with evaporation of the liquid on the specimen-bearing surface.

In some embodiments, the heater is positioned to deliver heat to the slide via the support element to produce a substantially uniform temperature profile along the portion of the specimen-bearing surface that bears the specimen. In some embodiments, the substantially uniform temperature profile has a temperature variation of less than 5% across the specimen-bearing portion of the specimen-bearing surface. In some embodiments, the substantially uniform temperature profile has a temperature variation of less than 4 ℃ across the specimen-bearing surface. Other temperature profiles can also be achieved.

In some embodiments, the heater comprises: at least two spaced elongate portions for conductively heating side portions of the support surface and two end heating portions of the support surface extending between the elongate portions. The two end heating portions are positioned to simultaneously heat a portion of the support surface for contacting one end of the slide and a portion of the support surface for contacting an area of the slide adjacent the slide label.

In some embodiments, the slide heating apparatus is configured to generate a low heating zone along a central region of the support surface and a high heating zone along the support surface. The high heating zone can surround (e.g., circumferentially surround) the low heating zone.

In some embodiments, the slide heating apparatus further includes a convection assembly positioned to generate a convection fluid through the recess defined by the heater to cool the support element. In some embodiments, the convection assembly includes one or more fans. The convective fluid is capable of cooling the support element without flowing through the specimen on the slide.

In some embodiments, the slide heating apparatus further comprises: a pair of side walls having a heat conductive portion and an insulating portion, respectively. The thermally conductive portion faces the slide to heat the slide.

In some embodiments, the slide heating apparatus further comprises: an overmolded retainer comprising an insulative material. Support elements are positioned between and supported by the sidewalls of the overmolded retainer. The thermal conductivity of the insulating material can be lower than the thermal conductivity of the material of the support element. In some embodiments, the insulating material comprises a non-metallic material (e.g., plastic) and the support element comprises metal.

In some embodiments, at least one of the heater and the support element comprises primarily stainless steel by weight. In some embodiments, the bearing surface comprises stainless steel. In some embodiments, a majority of the support element between the support element and the heater is stainless steel. The portion of the support element between the slide and the heater can have a thermal conductivity equal to or less than about 20W/m K.

In some embodiments, a method for heating a biological specimen carried on a slide includes: the slide is positioned on the support element of the conductive slide heating apparatus such that the back surface of the slide faces the support element and the specimen-bearing surface of the slide faces away from the support element. Heat can be delivered non-uniformly through the backside surface of the slide via the support element to substantially compensate for evaporative heat loss associated with evaporation of liquid on the specimen-bearing surface. Evaporative heat losses are non-uniformly across the specimen-bearing surface of the slide.

In some embodiments, a non-uniform temperature profile can be generated along the support surface of the support element that contacts the backside surface of the slide such that the specimen-bearing surface has a more uniform temperature profile than the non-uniform temperature profile. In some embodiments, the temperature change (e.g., the temperature change maintained in the portion of the specimen-bearing surface that contacts the biological specimen) can be equal to or less than about 5 ° while the temperature change of the support surfaces of the support elements that contact the backside surface of the slide exceeds 5 °.

The support surface of the support element can contact the backside surface of the slide and can be heated to create a low heating zone at a central region of the support surface and a high heating zone at a region of the support surface surrounding the central region. Additionally or alternatively, the support surface can be heated to create a high heating zone along the perimeter of the specimen-bearing surface along the staining region and a low heating zone at the central region of the staining region.

The slide can be conductively heated using thermal energy generated by a heating element of the conductive slide heating apparatus. The heating element comprises at least two spaced elongate heating portions and two end heating portions extending between the elongate heating portions. The elongate heating portion and the end heating portion define a convective cooling pocket for cooling the support element.

In some embodiments, a system for heating a slide bearing a specimen includes a slide platen including a support element, a conductive heater, and a controller. The support element has a support surface. The conductive heater is positioned to heat the support element. The controller is programmed to control the system to generate a non-uniform heating profile along the support element to transfer thermal energy to the slide to generate a substantially uniform temperature profile along the specimen-bearing area of the specimen-bearing surface of the slide when the back surface of the slide contacts the support surface.

In some embodiments, the conductive heater is configured to heat the support element to produce a non-uniform temperature heating profile across a majority of the support surface supporting the slide, thereby producing a substantially uniform temperature heating profile along a majority of the specimen-bearing surface of the slide. The substantially uniform temperature profile has a temperature change of less than 5 ° throughout the specimen-bearing region of the slide. Additionally or alternatively, the conductive heater can be configured to produce a low temperature heating zone along the support element and a peripheral high temperature heating zone along the support element. Additionally or alternatively, the conductive heater is located below the support element and defines an opening through which the convective fluid can cool the support element.

In some embodiments, a system for heating a slide bearing a specimen includes a convective cooling device coupled to a controller and configured to deliver a convective fluid into an opening based on a signal from the controller. In a particular embodiment, the convective cooling means comprises at least one fan capable of generating a convective fluid. In some embodiments, compressed air or motive air can be used.

In some embodiments, the support element comprises stainless steel. In some embodiments, a portion of the support element between the support surface for carrying the slide and the conductive heater has a thermal conductivity equal to or less than about 20W/m K.

Drawings

Non-limiting and non-exhaustive embodiments are described with reference to the following drawings. Unless otherwise specified, like reference numerals refer to like parts or acts throughout the various views.

FIG. 1 is an isometric view of a specimen processing system, in accordance with an embodiment of the disclosed technology.

Fig. 2 is an exploded isometric view of the specimen processing system of fig. 1. Portions of the protective housing are removed in the illustration.

FIG. 3 is a vertical projection view of a pipette device with a mixing station in accordance with an embodiment of the disclosed technology.

FIG. 4 is an isometric view of a carousel in accordance with embodiments of the disclosed technology.

FIG. 5 is a top plan view of the carousel of FIG. 4.

FIG. 6 is a cross-sectional view of the carousel taken along line 6-6 of FIG. 5.

FIG. 7 is a detailed view of a portion of the carousel of FIG. 6.

FIG. 8 is a bottom perspective view of a carousel in accordance with embodiments of the disclosed technology.

FIGS. 9A-9D illustrate various stages of operation of the pipette device.

FIG. 10 is a detailed view of a portion of the specimen processing system of FIG. 2.

Figure 11 is an isometric view of a slide ejector assembly in accordance with embodiments of the disclosed technology.

Fig. 12 is an isometric view of the slide ejector assembly of fig. 11 with the protective plate removed.

Fig. 13 and 14 are side views of the slide ejector assembly of fig. 11, with the slide carrier shown in different positions.

Fig. 15 is an isometric view of a slide staging device of the slide ejector assembly in which the slide is ready to be removed in accordance with an embodiment of the disclosed technology.

Fig. 16 is an isometric view of an empty sheet staging device in accordance with an embodiment of the disclosed technology.

Fig. 17 and 18 are top plan views of a slide staging device with an alignment device in accordance with embodiments of the disclosed technology.

Fig. 19 and 20 are isometric views of the slide ejector assembly with the protective plate removed.

Fig. 21 is a top plan view of the slide ejector assembly of fig. 19 and 20.

Fig. 22 is an isometric view of a slide staging device of a slide ejector assembly in accordance with another embodiment of the disclosed technology, wherein the slide is ready to be removed.

Fig. 23 is an isometric view of the slide staging device of fig. 22 illustrating components of an alignment device in accordance with an embodiment of the disclosed technology.

Fig. 24A and 24B are top plan views of a slide staging device with an alignment device in accordance with embodiments of the disclosed technology.

Fig. 24C and 24D are enlarged views of the alignment device of fig. 24B.

Fig. 25 and 26 are side views of a slide staging device and transfer assembly in accordance with embodiments of the disclosed technology.

Figure 27 is a block diagram illustrating a method for transferring a specimen slide using a specimen processing system in accordance with an embodiment of the disclosed technology.

Fig. 28 is an isometric view of an opposable dispenser, in accordance with embodiments of the disclosed technology.

Fig. 29 is a side view of the opposable dispenser of fig. 28.

Fig. 30 is an isometric view of a transport assembly and specimen processing station in accordance with an embodiment of the disclosed technology.

Fig. 31 is a side view of a transport assembly preparing delivery of opposable and slides to a specimen processing station in accordance with an embodiment of the disclosed technology.

Fig. 32 is a side view of an opposable actuator holding a opposable in accordance with embodiments of the disclosed technology.

Fig. 33 is an isometric view of a specimen processing station ready to process specimens on slides, in accordance with embodiments of the disclosed technology.

Figure 34A is a front isometric view, a top isometric view, a left isometric view of a slide holder platen holding a slide, in accordance with an embodiment of the disclosed technology.

Fig. 34B is a front isometric view, a top isometric view, a left isometric view of the slide holder platen of fig. 34A ready to hold a slide, in accordance with an embodiment of the disclosed technology.

Fig. 35 is a front isometric view, a bottom isometric view, a left isometric view of the slide holder platen of fig. 34A.

Fig. 36 is a bottom view of the slide holder platen of fig. 34A.

Fig. 37A is a cross-sectional isometric view of the slide holder platen taken along line 37A-37A of fig. 36.

Fig. 37B is a cross-sectional view of the slide holder platen taken along line 37B-37B of fig. 36.

Figure 38 is a top plan view of a specimen processing station holding specimen-bearing slides in accordance with an embodiment of the disclosed technology.

FIG. 39 is a cross-sectional view of a portion of the specimen processing station taken along line 39-39 of FIG. 38.

FIG. 40 is a cross-sectional view of a portion of the specimen processing station taken along line 40-40 of FIG. 38.

Fig. 41 is a cross-sectional view of the slide holder platen taken along line 41-41 of fig. 38.

Fig. 41A is a graph of position along a contact surface of a slide support versus thermal energy conducted to a slide, in accordance with an embodiment of the disclosed technology.

Fig. 41B is a graph of position along a contact surface of a slide support versus temperature of the contact surface in accordance with an embodiment of the disclosed technology.

Figure 41C is a graph of position along the upper surface of the slide versus temperature of the upper surface of the slide in accordance with an embodiment of the disclosed technology.

Fig. 42 is a top plan view of a heating zone created on a slide-supporting surface of a support element in accordance with an embodiment of the disclosed technology.

Figure 43 is a flow diagram illustrating a method for heating a slide in accordance with an embodiment of the disclosed technology.

Figure 44 illustrates a slide holder platen and dispenser assembly in accordance with embodiments of the disclosed technology.

Fig. 45 is a perspective view of a slide holder platen shown holding a slide in accordance with an embodiment of the disclosed technology.

Fig. 46 is a top view of the slide holder platen shown in fig. 45.

Fig. 47 is a perspective view of a slide holder platen according to the disclosed technology shown without a slide.

Fig. 48 is a partially exploded view of a slide holder platen.

Fig. 49 is an enlarged cross-sectional view of a portion of the slide holder platen shown in fig. 48.

FIG. 50 is a perspective view of a sealing member in accordance with an embodiment of the disclosed technology.

FIG. 51 is a cross-sectional end view of the seal member of FIG. 50 shown in an uncompressed configuration and a compressed configuration (shown in phantom).

Fig. 52A is a top view of the sealing member of fig. 50.

52B-52D are top views of sealing members according to various embodiments of the disclosed technology.

Fig. 53 is a cross-sectional side view of a portion of the slide holder platen prior to the slide engaging the sealing member.

Fig. 54 is a cross-sectional side view of a portion of the slide holder platen after a slide has been positioned on the slide holder platen.

Fig. 55 is an enlarged view of a portion of the slide holder platen shown in fig. 54.

Fig. 56 is an enlarged top view of a portion of the slide holder platen showing the sealing member in contact with the groove wall.

Figure 57 is a graph of equilibrium volume of liquid on a slide versus total evaporation rate of the liquid, in accordance with an embodiment of the disclosed technology.

FIG. 58 is a graph of time versus liquid coverage in accordance with an embodiment of the disclosed technology.

Fig. 59A and 59B are side and top views of a narrowed liquid band at one end of a gap between a opposable and a slide.

Fig. 60A and 60B are side and top views of a diffused liquid band.

Fig. 61A and 61B are side and top views of a liquid strip contacting a biological specimen.

Fig. 62A and 62B are side and top views of a liquid band between opposable and carrier adjacent regions of a label.

Fig. 63A and 63B are side and top views of a narrowed liquid band at one end of a gap adjacent a label of a slide.

Fig. 64 is an isometric view of a opposable according to one embodiment of the disclosed technology.

Fig. 65 is a top plan view of the opposable of fig. 64.

Fig. 66 is a side view of the opposable of fig. 64.

FIG. 67 is a detailed view of a portion of the opposable of FIG. 66.

Detailed Description

Fig. 1 shows a specimen processing system 100 ("system 100") that includes a protective housing 120, a slide carrier parking station 124 ("parking station 124"), an opposable carrier loading station 130 ("loading station 130"), and reagent parking stations 140, 142. The system 100 can automatically process specimen-bearing slides using opposables loaded via the loading station 130 to perform, for example, specimen conditioning (e.g., cell conditioning, washing, deparaffinizing, etc.), antigen retrieval, staining (e.g., H & E staining), or other types of protocols (e.g., immunohistochemistry protocol, in situ hybridization protocol, etc.) in order to prepare specimens for visual inspection, fluorescence visualization, microscopy, microscopic analysis, mass spectrometry, imaging (e.g., digital imaging), or other analytical or imaging methods. The system 100 can simultaneously process 20 specimen-bearing slides using the same or different protocols to provide processing flexibility and relatively high throughput. During processing (e.g., drying to staining), the specimen can be held on a slide for convenient handling and to prevent cross-contamination.

The protective housing 120 inhibits, limits, or substantially prevents contaminants from entering the internal processing environment. The protective housing 120 can include a cover 146 that can be opened to access internal components including, but not limited to, robotic components (e.g., robotic arms), transport devices (e.g., conveyors, actuators, etc.), fluidic components, specimen processing stations, slide platens, mixing components (e.g., mixing wells, reagent trays, etc.), slide carrier handling components, opposable carrier handling components, dryers, pressurizing devices (e.g., pumps, vacuums, etc.), and the like.

The parking station 124 includes an array of bay racks (bay). A slide carrier in the form of a basket is positioned in the left bay 148. Each bay can be configured to receive other types of slide carriers, such as racks, baskets, trays, or other types of carriers suitable for carrying slides before, during, or after specimen processing. The illustrated parking station 124 includes 12 bays separated by dividers. The number of bays, the location of the bays, the orientation of the bays, and the configuration of the bays can be selected based on the type of slide carrier to be used.

The loading station 130 comprises a receiving opening 150, through which receiving opening 150 a user can load the opposable carrier. The opposable carrier can be a cassette holding a stack of opposable elements. In other embodiments, the opposable carrier can be a cartridge or other portable structure for carrying the opposable.

The parking stations 140, 142 each include a row of bays. Each bay is capable of holding one or more containers, including bulk reagent containers (bulk reagent containers), bottles, bag-in-box (bag-in-box) reagent containers, and the like. The docking station 142 can hold a bulk liquid container that provides a liquid, such as a cleaning solution, for use in larger volumes. Empty containers in the docking stations 140, 142 can be conveniently replaced with full containers.

Movement of fluids into, out of, and within the specimen processing station can be controlled by fluidic modules including, for example, pumps, valves, and filters. The pneumatic module can supply pressurized air and generate vacuum to perform various slide processing operations and move fluids throughout the system 100. Waste can be delivered to the waste drawer 143. Figure 2 shows a waste drawer 143 holding waste containers 149A, 149B. The pneumatic module is capable of delivering waste material from the specimen processing station to the containers 149A, 149B, which containers 149A, 149B are capable of being periodically emptied.

The controller 144 can command system components and can generally include, but is not limited to, one or more computers, central processing units, processing devices, microprocessors, Digital Signal Processors (DSPs), Application Specific Integrated Circuits (ASICs), readers, and the like. To store information, the controller 144 can include, but is not limited to, one or more memory elements, such as volatile memory, non-volatile memory, Read Only Memory (ROM), Random Access Memory (RAM), and the like. The stored information can include: a heating program, an optimization program, a tissue preparation program, a calibration program, an indexing program, a blending program, or other executable program. Optimization procedures can be performed to optimize performance (e.g., enhance heating, reduce excess reagent consumption, increase productivity, enhance process consistency, etc.). The process can be optimized by determining, for example, an optimal arrangement to (1) increase the processing speed; (2) reducing the time of the heating or cooling cycle; (3) increasing throughput (e.g., increasing the number of slides processed in a particular length of time); and/or (4) reducing reagent waste. In some embodiments, the controller 144 determines a loading sequence for loading the specimen processing stations to reduce processing time and determine a loading sequence for the dispenser. This saves time because once a specimen-bearing slide is removed from the specimen processing station, fluid can be dispensed onto the next specimen-bearing slide. In some embodiments, the controller 144 determines a sequence for mixing and dispensing reagents using the mixing station 165.

Fig. 2 is an isometric exploded view of the specimen processing system 100, the specimen processing system 100 including the processing station 163, the slide ejector assembly 200, the opposable dispenser 380, and the specimen return mechanism 157. The processing station 163, slide ejector assembly 200, and opposable dispenser 380 are positioned at the left side of the internal environment 121. The specimen return mechanism 157 is positioned at the right side of the internal environment 121. The mixing station 165 is positioned generally below the specimen return mechanism 157 and can include a reservoir (e.g., a storage well). The reagents can be mixed in the mixing station 165. In other embodiments, the mixing station 165 can hold containers (e.g., vials, beakers, etc.) in which substances are stored and/or mixed. A row 152 of 20 specimen processing stations is capable of independently processing biological specimens.

In operation, a user can load slide carriers carrying slides bearing specimens into the empty bay of the parking station 124 of fig. 1 and can load opposable carriers carrying opposable items into the loading station 130. The slide carrier can be transferred to a reader (e.g., a label reader, a bar code reader, etc.) not shown that reads the label (if any) on the slide. The slide carrier can be delivered to the processing station 163, which can include, but is not limited to, a dryer (e.g., a dehydration unit), a heating unit (e.g., a drying module), or other components that can remove water from the slide, heat the specimen (e.g., heat the specimen to adhere the specimen to the slide), and the like. In some embodiments, the processing station 163 blows hot air over the slides to dry the slides, and if the specimen contains paraffin, the hot air can soften the paraffin to promote adhesion of the specimen to the slides. The air system can partially recirculate air to control humidity in the processing station 163. The slide carrier can be picked up and transported from the processing station 163 to another module (e.g., specimen processing station, label reader, etc.) or returned to one of the bays of the parking station 124.

The specimen return mechanism 157 is capable of loading slides bearing specimens into the slide carriers. The loaded slide carriers can be transported to the docking station 124. If the slide carrier is compatible with automatic coverslipping, the user can transport the slide carrier from the docking station 124 to an automatic coverslipper for coverslipping. Alternatively, slides can be manually coverslipped. Slides of the coverslip can be analyzed using an optical instrument (e.g., a microscope or other optical device).

FIG. 3 is a vertical projection view of a pipette device 172 in accordance with an embodiment of the disclosed technology. The pipette device 172 can be used as a staging area to provide improved staining characteristics, significantly improve throughput or otherwise enhance processing. The pipette device 172 is capable of preparing and holding large volumes of reagents (e.g., individual reagents and/or reagent mixtures). Particularly for reagents that react immediately upon mixing, the reactive reagents can be mixed immediately prior to dispensing to enhance dye consistency and quality. Because reagents can be staged well before they are needed, the pipette device 172 can improve slide processing capability and is well suited for use with large volume automated slide processing systems. In addition, the pipette device 172 can occupy a relatively small space and provide mixing and washing functions independent of slide processing.

In general, the pipette device 172 can include a mixing station 165, a reagent pipette assembly 175, and a wash pipette assembly 176. The mixing station 165 can include a carousel 177 and a drive mechanism 184 for rotating the carousel 177 about an axis of rotation 181. Carousel 177 can include a circular array of storage wells 180 (one identified) configured to hold large volumes of reagents. The drive mechanism 184 can rotate the carousel 177 (indicated by arrow 186) to position the reservoir wells 180 relative to the reagent pipette assembly 175 and/or the wash pipette assembly 176. The reagent pipette assembly 175 can partially or completely fill the reservoir wells 180 with fresh reagent from the filling station 209 (e.g., reagent shelf) and can also dispense reagent from the reservoir wells 180 onto a microscope slide. The reagent pipette assembly 175 may also be capable of washing and/or rinsing the reservoir wells or performing other operations. The wash pipette assembly 176 is capable of washing the reservoir well 180 by, for example, flushing the reservoir well 180 with a wash solution and vacuuming a liquid (e.g., wash solution, reagents, etc.) from the reservoir well 180. Fresh reagents can be mixed in the purged storage well 180.

FIG. 4 is a front top isometric view of carousel 177 in accordance with embodiments of the disclosed technology. Fig. 5 is a top plan view of the carousel 177. Referring to fig. 4 and 5 together, carousel 177 can include storage wells 180 (one identified), ramps 182, and discharge tubes 183. The storage wells 180 can be angularly spaced (uniformly or non-uniformly) about the discharge tube 183, and each storage well 180 can contain a sufficient volume of liquid for one or more dispensing steps in a staining protocol. In some embodiments, each storage well 180 has a containment capacity in the range of about 200 μ L to about 450 μ L. In one embodiment, each storage well 180 has a containment capacity of approximately 350 μ L. In other embodiments, different storage wells 180 can have different holding capacities to prepare different volumes of reagent mixtures. The holding capacity of the reservoir well 180 can be selected based on the desired volume of reagent mixture to be dispensed. A set of storage wells 180 (e.g., four storage wells) can correspond to a particular slide and/or slide processing station to prevent cross-contamination. In a staining protocol that utilizes a quantity of reagent mixture, a reservoir well (e.g., adjacent reservoir well 180) can be used to prepare and contain the reagent mixture. In some embodiments, carousel 177 can include multiple arrays of wells positioned at different locations relative to discharge tube 183. For example, multiple circular arrays of storage wells can be positioned at a different radius than the central discharge pipe radius of the central discharge pipe 183.

The reservoir well 180 can be oriented in a generally vertical orientation (e.g., the longitudinal axis of the reservoir well can be oriented vertically) to access the bottom of the reservoir well 180 using a vertically oriented pipette. Reservoir well 180 may be circular (fig. 5), oval, elliptical, combinations thereof, or other shapes without sharp corners to facilitate flushing/cleaning. The illustrated carousel 177 has a plurality of storage wells 180 (e.g., forty storage wells 180) to allow for rapid processing of a relatively large number of slides (e.g., up to about a hundred slides or more), but the carousel 177 can have a greater or lesser number of storage wells 180 to increase or decrease the number of slides served by the carousel 177. The geometry (e.g., circular, elliptical, etc.), mode (e.g., circular array, elliptical array, etc.), number, and orientation of the reservoir wells 180 can be selected based on the number of slides, staining protocol, and operation of the reagent pipette assembly 175 and/or the wash pipette assembly 176.

A ramp 182 can extend between the storage well 180 and the discharge pipe 183. Spilled liquid (e.g., reagents, cleaning fluids, or mixtures thereof) exiting the storage well 180 can flow along the upper surface 185 of the ramp 182 and through the drain 183. In some embodiments, the upper surface 185 slopes downward toward the drain 183 and has a shape (e.g., a generally frustoconical shape) for promoting radially inward flow. The upper surface 185 can help keep the streams from two or more storage wells 180 separate to inhibit or limit mixing of the streams to avoid or mitigate unintended chemical reactions. In some embodiments, the ramp 182 has a channel, groove, or other feature that facilitates the flow of spilled liquid toward the drain 183.

Referring now to fig. 4, the carousel 177 can include spillways 187 (one identified) configured to allow overflow liquid to be automatically drained from the storage wells 180. Spillway 187 can prevent cross contamination by preventing well-to-well flooding. During a cleaning cycle, the storage wells 180 can be filled with a cleaning solution (e.g., water, deionized water, cleaning solution, etc.) without affecting adjacent storage wells 180. In some embodiments, spillway 187 includes spill partition 189 (two identified in fig. 4 and 5) and spill wall 190. Each spacer 189 can be positioned between adjacent storage wells 180.

FIG. 6 is a cross-sectional view of carousel 177 taken along line 6-6 of FIG. 5. Fig. 7 is a detailed view of a portion of carousel 177. Referring now to fig. 7, a partition 189 can prevent splashed liquid from reaching a nearby storage well and can include an outer portion 192 and an inner portion 194. In some embodiments, the spacer 189 can be positioned between the center of adjacent storage wells 180 and other storage wells (e.g., 1/5, 1/4, or 1/3 for the total number of storage wells 180). During a cleaning cycle, the cleaning fluid can tend to spray and/or splash, and the spacer 189 can block such spray/splash, thereby preventing cross-contamination between wells. The size and configuration of the spacers 189 can be selected to keep the storage wells fluidly isolated from each other.

The outer portion 192 can be positioned directly between two storage wells and can extend upwardly through a spillway inlet in the form of an edge 196 of the wall 190. In some embodiments, outer portion 192 extends upwardly past edge 196 a sufficient distance to prevent well-to-well flooding. For example, the height H of the outer portion 192 can be in the range of about 3 mm to about 7 mm. Other heights can be used if needed or desired. The interior 194 can be a generally vertically oriented wall that extends inwardly (e.g., toward the center of the carousel 177). The length 199 of the interior 194 can be approximately equal to the height H to prevent directing liquid (e.g., wash or reagent) toward an unintended wall at risk of cross-contamination. The length L of the spacer 189 can be equal to or greater than the diameter D of the reservoir well 180. For example, the ratio of the length L to the diameter D can be equal to or greater than 1.25, 1.5, 2, or 2.5.

The storage well 180 has a generally smooth sidewall 193 (e.g., a cylindrical sidewall or other shaped sidewall without sharp corners) and a bottom 195 (fig. 6), the bottom 195 defining a cavity capable of accommodating a desired volume, e.g., 250 µ L, 350 µ L, or 450 µ L. Fig. 7 shows a liquid line 198 (illustrated in dashed lines) of the desired volume of reagent. When excess liquid is delivered to the storage well 180, the liquid can rise above the inlet 196 of the spillway 180 and cause an overflow. As shown in fig. 7, liquid 201 (illustrated in phantom) can overflow wall 190 and flow along upper surface 185. Referring now to fig. 6, liquid 201 can exit carousel 177 via a drain 183, which drain 183 can be large enough to contain fluids discharged from multiple storage wells. Flooding can happen intentionally to flush the storage wells, and flooding can happen accidentally, for example, if excess reagent is dispensed into one of the storage wells.

Fig. 7 shows stops 313 (one identified) that limit the maximum plunge depth of the pipette to prevent damage to carousel 177 that may be caused by, for example, over-insertion of the pipette. The stops 313 can be circumferentially spaced from one another and can extend upwardly a sufficient distance 315 to prevent the wash pipette 213 and/or the reagent pipette 204 from contacting the bottom 195 of the reservoir well. For example, a head assembly carrying a pipette can hit the stop 313 before the pipette carried by the head assembly damages the carousel 177. Other types of stops can be used to position or limit the movement of the pipette.

FIG. 8 is a bottom perspective view of the carousel 177 including mounting bayonets 205 and alignment features 207. The mounting bayonet 205 can be coupled to a drive shaft of a drive mechanism (e.g., the drive mechanism 184 of fig. 4) and can include one or more locators 218. In other embodiments, the outer surface of the carousel 177 can be used to rotate the carousel 177. For example, the drive wheel can engage an outer surface of the carousel 177 such that rotation of the drive wheel causes rotation of the carousel 177. The detents 218 can be flanges, ribs, or other features that are matable with the drive shaft of the drive mechanism. The alignment features 207 can be used to visually, mechanically, electromechanically, and/or optomechanically align the carousel 177. In some embodiments, the alignment feature 207 is a notch or cutout that receives an alignment protrusion of the drive mechanism. In other embodiments, the alignment features 207 can be protrusions or other visually (including optically) identifiable features for conveniently identifying and orienting the carousel 177. In some embodiments, the alignment features 207 can be used to register the carousel 177 so that the location of individual storage wells 180 is known to the control system (e.g., controller 144). The upper edge or surface 231 can be located a critical distance from the bottom of the skirt 235 in which it resides such that if the sensor (e.g., optical sensor) does not recognize the alignment feature 207, the user will be immediately notified that the carousel 177 is improperly installed. The carousel described herein can be conveniently removed from the drive mechanism 184 to clean it or replace it, and the alignment features 207 can be used to reinstall the carousel 177 onto the drive mechanism 164. One side of the alignment feature 207 can be detected and used to inform an operator whether the carousel 177 is improperly installed.

The single piece carousel can have a unitary construction and can be formed by a molding process, a machining process, or other suitable process. For example, the carousel 177 can be monolithically formed by an injection molding process. In a multi-piece embodiment, the carousel 177 can have a carousel body and separate spillway and storage wells mounted in the carousel body. The configuration of the carousel 177 can be selected based on the desired functionality of the carousel 177.

Fig. 9A-9D illustrate the operation of the pipette device 172. Generally, the reagent pipette assembly 175 can sequentially deliver fresh reagents to the storage wells 180 to create a reagent mixture. The reagent pipette assembly 175 can deliver such reagent mixtures onto the slide at the slide processing station. The carousel 177 can be rotated to sequentially position the reservoir wells 180 at a cleaning position for cleaning by cleaning the pipette assembly 176. In some embodiments, the reagent pipette assembly 175 can mix reagents while the wash pipette assembly 176 washes the reservoir wells 180 to reduce overall processing time. In other embodiments, reagent mixing and reservoir well cleaning are performed at different times. The pipette cleaner 251 can purge (e.g., using a cleaning solution), vacuum draw, air blast purge, or otherwise clean the pipette 204 between each stroke to the filling station 209 to prevent cross-contamination of reagents. The pipette cleaner 251 can also clean the pipette 213 between wash operations. The operation of the reagent pipette assembly 175, the wash pipette assembly 176, and the mixing station 165 is described in detail below.

FIGS. 9A-9C illustrate one method of using the reagent pipette assembly 175. The reagent pipette assembly 175 can have different types of pipettes, valves, and sensors, and in some embodiments can be similar or identical to the pipette dispensers 160, 162 depicted in fig. 2. In various embodiments, the reagent pipette assembly 175 can include a positioning mechanism having one or more track/carriage assemblies, motors (e.g., drive motors, stepper motors, etc.), drive elements (e.g., chains, belts, etc.), or other features for providing motion. The reagent pipette assembly 175 can take fresh reagents, stage the reagents and dispense the reagents onto a microscope slide. In some embodiments, the reagent pipette assembly 175 can move the reagent pipette 204 to, for example, a filling position at the filling station 209 (see fig. 9A), an unloading/loading position for dispensing reagent into one of the reservoir wells 180 or loading the pipette 204 with reagent from one of the reservoir wells (fig. 9B), and a dispensing position for dispensing reagent onto a slide at the slide processing system (fig. 9C).

Referring now to fig. 9A, the reagent pipette assembly 175, in the reagent loaded operating state, can insert the pipette 204 into one of the containers 211 at the filling station 209 and can draw a desired volume of fresh reagent 227. In some embodiments, the reagent pipette assembly 175 is capable of drawing a vacuum provided by the pressurizing device 221. Pressurizing device 221 can include one or more vacuum sources, pumps, or other devices capable of providing a desired vacuum or positive pressure. The container 211 can be, but is not limited to, a vial, a bottle, a jar, or other container suitable for containing a substance for processing a specimen. The illustrated filling station 209 has three containers 211, but a greater or lesser number of containers can be used, and the filling station 209 can be part of a docking station, such as docking stations 140, 142 of FIG. 1. For example, the containers 211 can be mounted in bays of the docking stations 140, 142 of fig. 1 and can be accessed by a reagent pipette assembly 175, which reagent pipette assembly 175 can move through the internal environment 121 of fig. 2.

Fig. 9B shows the reagent pipette assembly 175 after the reagent pipette 204 has been filled with reagent. The pipette 204 is positioned to deliver the reagent into the reservoir well 180 identified in fig. 9B. Pressurizing device 221 can provide positive pressure to dispense reagents. The reagent pipette assembly 175 can take additional reagent from the filling station 209 and dispense it into the same reservoir well 180 to produce a reagent mixture.

Referring to fig. 9B and 9C, to dispense the reagent mixture held by carousel 177, reagent pipette 204 can be inserted into reagent well 180 and filled with a desired volume of reagent mixture. Fig. 9C shows the loaded reagent pipette 204 dispensing the reagent mixture onto the microscope slide 156 at the processing station 245. The reagent pipette assembly 175 can repeatedly obtain reagents from the mixing station 165 and dispense the reagents onto the slide 156 or other slides at other processing stations.

Fig. 9C and 9D illustrate stages of a washing process performed by washing the pipette assembly 176. Generally, the storage wells 180 can be cleaned by, for example, dispensing a cleaning fluid to fill the storage wells 180 and removing (e.g., aspirating) the cleaning fluid and any residual reagents remaining in the storage wells 180. The wash pipette assembly 176 can include a vacuum source 237 and a pressurizing device 239 connected to the wash head assembly 241 by conduits 247, 249, respectively. The drive assembly 184 is capable of rotating the carousel 177 to position the reservoir well 180 at a wash position below the wash pipette 233.

Fig. 9D shows the wash pipette 233 after the wash pipette 233 has been lowered into one of the reservoir wells. The cleaning fluid can be delivered through the cleaning pipette 213 to dilute the reagents (if any) in the reservoir wells, flush the reservoir wells, and/or otherwise rinse or clean the reservoir wells. In some embodiments, vacuum source 237 can be activated and wash pipette 213 can aspirate most or substantially all of the reagent in reservoir well 180. The storage well 180 can then be filled with a cleaning solution that flows (indicated by arrows) in a controlled manner to the drain 183. The overflow process can remove most or substantially all of the residual reagent volume within the storage well 180. After flushing the storage well 180, the vacuum source 237 can be activated again to clean the storage well. In other embodiments, prior to pumping, the storage well can be flooded with a cleaning solution that flows (indicated by arrows) in a controlled manner to the drain 183. The flooding process can remove most or substantially all of the reagent volume in the storage well. After flushing the reservoir well, the vacuum source 237 can be activated, and the wash pipette 213 can aspirate most or substantially all of the liquid (e.g., wash solution, a mixture of wash solution and reagents, etc.) remaining in the reservoir well 180. Then, the pipette 213 can be raised and the drive mechanism 184 can rotate the carousel 177 to position another reservoir well at a wash position (e.g., wash beneath the pipette 213). The pipette cleaner 251 (fig. 9A) can periodically clean the outside of the pipette 213. In other embodiments, two or more pipettes can be used during the wash. For example, one wash pipette can be used to dispense wash liquid and another wash pipette can aspirate residual liquid from the reservoir well. In yet another embodiment, the reagent pipette assembly 175 can be used to perform a wash cycle by flushing out the reservoir well 180.

The controller 144 of fig. 9D can be configured to command the drive mechanism 184 to sequentially move each of the reservoir wells 180 to a wash position for washing by washing the pipette assembly 176. In some embodiments, the controller 144 stores instructions in the memory 147 (illustrated in phantom) and executes the instructions to command the pipette device 172 to sequentially fill the reservoir well 180 with reagents from the containers 211. Additionally or alternatively, the memory 147 can store mixing instructions (e.g., a mixing program) that are executable by the controller 144 to command the wash pipette assembly 176 to deliver at least two reagents (e.g., two reagents, three reagents, etc.) to one of the storage wells. The mixing instructions can be selected based on information obtained from the slide to be processed. The controller 144 can be communicatively coupled to any or all of the components of the pipette device 172.

The system 100 of fig. 1 and 2 can include one or more pipetting devices 172 as discussed in connection with fig. 3-9D. The system 100 can have mixing stations 165 at opposite sides of the internal environment 121 (fig. 2). The wash pipette assembly can be stationary and the wash pipette vertically movable to avoid collisions between the wash pipette and a reagent pipette that can move around the mixing station. The mixing station 165 can be serviced by a single reagent pipette assembly and a single wash pipette assembly. In other embodiments, each mixing station 165 is serviced by a respective reagent pipette assembly and wash pipette assembly. The number of mixing stations, the location of the mixing stations, and the sequence of operation of the reagent and wash pipette assemblies can be selected based on the procedure to be performed.

Fig. 10 is a detailed view of a segment of row 152. The opposable element 154 ("opposable 154") is capable of moving a substance along the slide 156 to contact a specimen on the slide 156. In some embodiments, including the illustrated embodiment, a series of substances can be used to independently process 20 slides.

If the specimen is a biological sample embedded in paraffin, the sample can be dewaxed using an appropriate dewaxing fluid (or fluids). After removing the dewaxing fluid (or fluids), any number of substances can be successively applied to the specimen using opposable 154. Fluids can also be applied for pretreatment (e.g., protein cross-linking, exposing nucleic acids, etc.), denaturation, hybridization, washing (e.g., stringency washing), detection (e.g., coupling a visual or marker molecule to a probe), amplification (e.g., amplifying a protein, gene, etc.), counterstaining, etc. In various embodiments, the substance includes, but is not limited to, a stain (e.g., hematoxylin solution, eosin solution, etc.), a wetting agent, a probe, an antibody (e.g., monoclonal antibody, polyclonal antibody, etc.), an antigen retrieval fluid (e.g., an aqueous or non-aqueous based antigen retrieval solution, antigen retrieval buffer, etc.), a solvent (e.g., alcohol, limonene, etc.), and the like. Stains include, but are not limited to, dyes, hematoxylin stains, eosin stains, conjugates of antibodies or nucleic acids with detectable labels (e.g., haptens, enzymes, or fluorescent moieties, etc.), or other types of substances for imparting color and/or for enhancing contrast. In some embodiments, the substance applied is a liquid reagent applied via a dispenser, such as pipette dispensers 160, 162 depicted in fig. 2 or reagent pipette assembly 175 depicted in fig. 3-9D.

The biological specimen can include one or more biological samples. The biological sample can be one or more tissue samples (e.g., a collection of any cells) removed from a subject. The tissue sample can be a collection of interconnected cells that perform similar functions within an organism. The biological sample can also be any solid or fluid sample obtained from, excreted by, or secreted by any living organism, including but not limited to: unicellular organisms such as bacteria, yeast, protozoa and amoebae; multicellular organisms (e.g., plants or animals, including samples from healthy or superficially healthy human subjects or human patients afflicted with a condition or disease to be diagnosed or investigated, such as cancer). In some embodiments, the biological sample is mountable on a microscope slide and includes, but is not limited to, a section of tissue, an organ, a tumor section, a smear, a frozen section, a cytological preparation (cytology prep), or a cell line. Excisional biopsy, core biopsy, excisional biopsy, needle biopsy, core needle biopsy, stereotactic biopsy, open biopsy or surgical biopsy can be used to obtain the sample.

Fig. 10 shows a rack carrying a set of sealed containers 211, the containers 211 each containing from about 10 mL to about 30 mL of reagent. The sealed container 211 has a cap 151 with a sealing element in the form of a membrane 153 that can minimize, limit, or substantially prevent evaporative loss. The septum 153 can be broken (e.g., punctured, torn, etc.) to access the contents of the container 211. When the user installs the container 211, the membrane 153 can be broken to establish fluid communication with a pump or pipette (e.g., reagent pipette 204 of fig. 9A-9D), which in turn delivers the fluid to the appropriate specimen processing station. The container 211 can include, but is not limited to, one or more human-readable labels, machine-readable labels (e.g., bar codes to be read by the system 100), or other types of labels. In some embodiments, the docking station 140 provides fluids and solutions (e.g., dye solutions, such as hematoxylin solution and eosin solution) that are used in smaller volumes.

Fig. 11 and 12 show a slide carrier 170 loaded into a slide ejector assembly 200 ("ejector assembly 200"). The plate 216 of fig. 11 is removed in the illustration of fig. 12. The ejector assembly 200 includes a slide carrier handler 202 ("carrier handler 202"), a slide staging device 210 ("staging device 210"), and an ejector 212. Carrier handler 202 can include a carrier receiver 220 (fig. 12) and a receiver rotation device 224 (fig. 12). The carrier receiver 220 includes a pair of spaced apart arms 226 (e.g., elongated members, cantilevered members, etc.) on which the slide carrier 170 can rest. The illustrated slide carrier 170 is a slide rack capable of holding microscope slides in a spaced arrangement. One slide is shown in carrier 170 of fig. 11 and 12. In some embodiments, the slide carrier 170 can be baskets, such as SAKURA baskets or similar baskets with shelves or partitions.

Carrier receiver 220 of fig. 12 can include one or more grippers, clamps, holders, or other components that releasably hold slide carriers. The receiver rotation device 224 can include, but is not limited to, one or more motors, actuators, or other components capable of rotating the arm 226. The arm 226 can move along an arcuate rail, pivot mechanism, or the like to rotate the slide carrier 170. The carrier handler 202 can also include a carriage 230 and a track 232. The carriage 230 can travel along a track 232 to move the slide carrier 170 vertically.

Referring again to fig. 11, a fully or partially loaded slide carrier can be inserted between the plates 214, 216. The receiver rotation device 224 (fig. 12) is capable of rotating the carrier receiver 220 from the loading position 213 (fig. 11) holding the slide in a substantially vertical orientation to the intermediate position 215 (fig. 13) holding the slide in a substantially horizontal orientation. The term "substantially horizontal" generally refers to an angle within about +/-3 degrees of horizontal, e.g., within about +/-1 degree of horizontal, e.g., within about +/-0.8 degrees of horizontal. The slide carrier 170 can be moved vertically to an unload position 217 (fig. 14). The ejector 212 is capable of sequentially moving the specimen-bearing slides to the staging device 210. The staging device 210 is capable of positioning the specimen-bearing slide for subsequent transport, as discussed in connection with fig. 15-18.

Fig. 15 and 16 are isometric views of staging device 210 including a standby platform 240 and an alignment device 242. The standby platform 240 can include a cantilever plate 248, a slide holding region 250 ("holding region 250"), and an over travel inhibitor 254. In fig. 15, the slide 243 rests on a holding region 250, which holding region 250 can be a raised area that is smaller than the slide 243. The slide 243 can protrude outward from the holding region 250 such that excess fluid (if any) can drain from the slide 243 onto the plate 248 without capillary action beneath the slide 243 (e.g., between the slide 243 and the surface 361 of fig. 16). In some embodiments, the standby platform 240 can include, but is not limited to, one or more sensors, readers, heaters, dryers, or other components that facilitate processing of slides.

Referring to fig. 16, the over travel inhibitor 254 is capable of accurately positioning the slide without physically contacting the slide, the edges of the label, and/or a specimen on other areas of the slide that may affect positioning accuracy. In some embodiments, the over travel inhibitor 254 is capable of positioning the slide without contacting the top of the slide at a location, for example, near a hang tag, which can affect positioning accuracy. The over travel inhibitor 254 includes a vacuum port 290 and a vacuum source 281, the vacuum source 281 being fluidly coupled to the vacuum port 290 via one or more fluid conduits 283 (e.g., an inner fluid conduit, an outer fluid conduit, etc.). Vacuum source 281 can include, but is not limited to, one or more pressurization devices, pumps, or other types of devices capable of drawing a vacuum through opening 310. The bottom surface of the slide 243 (fig. 15) and the contact surface 300 of the vacuum port 290 can form a seal to maintain the vacuum. In some embodiments, the contact surface 300 can include one or more compressible materials (e.g., rubber, silicon, etc.) capable of maintaining a hermetic seal. In other embodiments, the contact surface 300 can comprise one or more non-compressible materials (e.g., aluminum, stainless steel, etc.), and in some embodiments, can comprise one or more sealing members (e.g., O-rings, gaskets, sealing cups, etc.) for forming a seal with the slide 243. In further embodiments, contact surface 300 and/or vacuum port 290 can include a pressure sensor or other sensor for detecting the presence of blade 243 on standby platform 240.

The retention region 250 includes ends 320, 322 and a body 328 extending between the ends 320, 322. The ejector stop 314 is defined by the end 320 and can be used to reference the position of one end of the slide 243. The ejector stop 314 can be a sidewall or edge of the end 320. In other embodiments, the ejector stop can be one or more protrusions.

As shown in the embodiment illustrated in fig. 16-18, staging device 210 includes an alignment device 242. In one embodiment, the alignment device 242 includes a pair of generally parallel clips 270, 272 that project upwardly through the openings 277, 279, respectively, and vertically through the retention region 250, the clips 270, 272. The alignment device 242 can include, but is not limited to, one or more actuators (e.g., pneumatic actuators, electromechanical actuators, etc.) capable of moving the grippers 270, 272. Because the transfer head may not be able to properly pick up and handle a misaligned slide, the alignment device 242 is able to align the slide to facilitate the pick up and handling of the slide. In some embodiments, the label of the slide can be spaced from the holders 270, 272 to prevent the slide from undesirably adhering to the holders 270, 272.

Fig. 17 shows the longitudinal axis 271 of the slide 243 in a misaligned position. The longitudinal axis 271 is non-parallel with the longitudinal axis 273 of the holding region 250. The jaws 270, 272 can be moved (indicated by arrows 280, 282) toward each other from an open position (fig. 17) to a closed position (fig. 18) to reposition the slide 243. In some embodiments, the longitudinal axis 271 of the slide 243 in the aligned position can be substantially aligned with (e.g., parallel to) the longitudinal axis 273 of the holding region 250. After the slide 243 is aligned, the grippers 270, 272 can be returned to the open position and the now aligned slide 243 can be picked up. The configuration and operation of the alignment device 242 can be selected based on the desired position of the aligned slide. In addition, because the grippers 270, 272 apply the same force to opposite sides of the slide, the alignment device 242 can be used to align slides having different sizes.

Fig. 19-21 illustrate an ejector 212 that includes an ejector member 330, a base 334, and a drive mechanism 336. The ejector element 330 includes an elongated portion 340 located in a recess 341 in the base 334 and a mounting portion 342 coupled to a rod 344 of the drive mechanism 336. The drive mechanism 336 can provide reciprocating linear motion and can include, but is not limited to, one or more stepper motors, pistons (e.g., pneumatic pistons, hydraulic pistons, etc.), pressurizing devices (e.g., pumps, air compressors, etc.), sensors, and the like. The illustrated rod 344 has been moved in the direction indicated by arrow 350 to move the ejector element 330 from a first or initial position 351 (illustrated in phantom in fig. 21) through the slide carrier receiving gap 352 ("gap 352") such that the head 360 of the elongate portion 340 pushes the slide onto the standby platform 240. The head 360 can include a compliant material (e.g., rubber, plastic, etc.) to avoid damaging the slide. In some embodiments, the head 360 can push the slide along the surface 361 (fig. 16) of the holding region 250 until the slide is at a desired position. Slides can be removed from the slide carrier 170 one at a time until the slide carrier 170 is empty.

Referring again to fig. 1 and 2, a user can load a slide carrier holding slides bearing specimens into the docking station 124. The transfer mechanism is capable of transporting the slide carrier to the ejector assembly 200. The transfer mechanism can include, but is not limited to, one or more robotic arms or arms, an X-Y-Z transport system, a conveyor, or other automated mechanism capable of transporting items between locations. In some embodiments, the transfer mechanism includes one or more end effectors, grippers, suction devices, holders, clamps, or other components suitable for grasping slide carriers.

The ejector assembly 200 moves the slide carrier 170 to the unload position 217 (fig. 14). The slide carrier 170 is moved vertically to index the slide relative to a reference position. The reference position can be a plane defining a slide removal position (e.g., the fixed slide removal plane 275 shown in fig. 14). The bottom of the slide to be removed can be substantially coplanar, or slightly above the surface 361 (fig. 16). The drive mechanism 336 can move the ejector element 330 horizontally to move the elongated portion 340 (fig. 19) through the carrier 170 to push the slide onto the surface 361 (fig. 15). When the head 360 contacts the ejector stop 314 (fig. 16), a vacuum can be drawn through the slide over travel inhibitor 254 to inhibit movement of the slide 243. The head 360 can then be removed from the slide 243. The grippers 270, 272 can be moved from an open position to a closed position to align the slide 243. The aligned slide 243 can be retrieved and transported to the specimen processing station. The drive mechanism 336 can move the ejector element 330 back and forth and the slides can be indexed to sequentially deliver all of the slides to the staging device 210.

To protect the specimen, the lowermost slide in the slide carrier 170 can be ejected first. By starting from the lowermost slide, the specimen (or specimens) on the vertically adjacent slide can face away from the head 360 and thus be protected. If the head 360 is not vertically aligned with the slide to be removed, the head 360 may hit the bottom of the vertically adjacent slide without dislodging the specimen (or specimens) on the upper surface of the vertically adjacent slide. After removing the lowermost slide, the lowermost slide remaining in the slide carrier 170 can be removed. This process can be repeated until the slide carrier 170 is empty. Other indexing sequences can be used to remove slides.

The empty slide carrier 170 can be returned to the loading position (fig. 11) and subsequently transported to one of the bays of the parking station 124. The empty slide carrier 170 can be removed from the docking station 124 and filled with slides bearing specimens and returned to the docking station 124. Alternatively, an empty slide carrier 170 can be filled with processed specimen-bearing slides using the ejector assembly 200. The pusher assembly can be used to push processed specimen-bearing slides on the staging device 210 into a slide carrier. Thus, the ejector assembly 200 can be used for both unloading and loading slide carriers.

Fig. 22-26 illustrate a staging device 210a of a slide ejector assembly 200a configured in accordance with another embodiment of the present technique. Fig. 22 and 23 are isometric views of a staging device 210a, the staging device 210a including features that are substantially similar to the features of the staging device 210 described above with reference to fig. 16-18. For example, staging device 210a includes a staging platform 240a having a cantilevered plate 248a (similar to staging platform 240 shown in fig. 16), a slide holding region 250a ("holding region 250 a"), and an over-travel inhibitor 254a (similar to over-travel inhibitor 254 shown in fig. 16). The staging device 210a also includes an alignment device 242a configured to move the slide 243 from a misaligned position to an aligned position on the standby platform 240 a. However, in the embodiment shown in fig. 22 and 23, the alignment device 242a does not include a pair of generally parallel clamps 270, 272 (fig. 16) that project upwardly through openings 277, 279 (fig. 16) in the staging platform 240 a.

In the embodiment illustrated in fig. 22, the alignment device 242a includes a first alignment member 362 for engaging the first edge 244 of the slide 243 and a second alignment member 364 positioned opposite the first alignment member 362 for engaging the second edge 245 of the slide 243. The engagement of the first side 244 and the second side 245 of the slide 243 can cause the slide 243 to pivot or otherwise move from a misaligned orientation on the slide holding region 250a to an aligned orientation on the holding region 250a to facilitate slide pickup and handling by a transfer apparatus (not shown).

Referring to fig. 23, first and second alignment members 362, 364 are secured to blocks 365, 366 by first and second fasteners 367, 368 (e.g., pins, bolts, screws, or other mechanical fasteners known to those skilled in the art). For example, the blocks 365, 366 can include holes 369, 370 for receiving fasteners 367, 368, respectively. The blocks 365, 366 can also include one or more protrusions 371, 372 for allowing rotation or pivoting of the alignment members 362, 364 and for engaging the first and second alignment members 362, 364, respectively, to limit rotation or pivoting of the alignment members 362, 364 relative to the blocks 365, 366 and/or during engagement with the slide 243 (described below). Openings 373, 374 (one identified) can be provided in the alignment members 362, 364 for receiving the protrusions 371, 372. In other embodiments, the protrusions may be provided on the alignment members 362, 364, which alignment members 362, 364 may be received in openings provided in the blocks 365, 366. In some embodiments, the protrusions 371, 372 may be non-circular with a rectangular or other geometric shape. The openings 373, 374 can be shaped to accommodate the respective geometry of the protrusions 371, 372, or, as illustrated in fig. 23, the openings 373, 374 can be through-holes that receive the protrusions 371, 372.

The alignment device 242a can include, but is not limited to, one or more actuators (e.g., pneumatic actuators, electromechanical actuators, etc.) that can move the blocks 365, 366 having the alignment members 362, 364 secured to the blocks 365, 366 toward and away from the longitudinal axis 273a of the holding region 250a (shown in fig. 24A and 24B). For example, fig. 24A and 24B are enlarged top views of the staging device 210a illustrating stages in the process for aligning the longitudinal axis 271a of the slide 243 with the longitudinal axis 273a of the holding region 250 a. Figure 24A shows the longitudinal axis 271a of the slide 243 in a misaligned position. The longitudinal axis 271a is non-parallel to the longitudinal axis 273a of the holding region 250 a. The first and second alignment members 362, 364 are movable toward one another (indicated by arrows 375, 376) from an open position (fig. 24A) to a closed position (fig. 24B) in which the alignment members 362, 364 engage or contact the first and second sides 244, 245 of the slide 243 to reposition the slide.

In one embodiment, the first and second alignment members 362 and 364 together contact the slide 243 at three separate contact points. In the embodiment illustrated in fig. 24B and 24C, the first alignment member 362 has a first contact region 377 and a second contact region 378 configured to engage the first edge 244 of the slide 243. As illustrated in fig. 24B and 24D, the second alignment member 364 has a third contact region 379 configured to engage the second edge 245 of the slide 243. In one embodiment, the area of contact points is the portion of the carrier sheet 243 to which the first, second, and third contact regions 377, 378, and 379 engage. In some arrangements, the contact points are relatively small discrete portions of the slide 243 (e.g., along the first and second edges 244, 245). In some embodiments, the surface areas defined by the three contact points and engaged by the first 377, second 378, and third 379 contact regions are substantially the same; however, in other embodiments, the surface area can be different. In one embodiment, the third contact region 379 is configured to contact the second edge 245 of the slide 243 at a lateral position along the slide 243 between the lateral position contacted by the first contact region 377 and the second contact region 378 on the first edge 244 of the slide 243.

Referring to fig. 24B, the slide 243 is able to move (e.g., pivot about a midpoint or axis of rotation 246 created or defined by three separate points of contact) to an aligned position when the first and second contact regions 377 and 378 of the first alignment member 362 and the third contact region 379 of the second alignment member 364 engage the first and second sides 244 and 245 of the slide 243, respectively. The movement of the first and second alignment members 362, 364 through blocks 365, 366 can continue until the slide 243 is engaged by the first, second, and third contact regions 377, 378, and 379, and the slide 243 is no longer moving (e.g., resting on the holding region 250a in an aligned position). In some embodiments, the first and second alignment members 362, 364 may include one or more pressure sensors 381 (fig. 24C and 24D) on or adjacent to one or more contact regions 377, 378, 379 to ensure that the alignment members 362, 364 apply a force of sufficient magnitude to move the slide 243 and/or do not compress the slide 243 in a manner that may damage or compromise the slide. In some embodiments, the contact regions 377, 378, 379 can include coatings and/or compliant materials (e.g., rubber, plastic, etc.) to avoid damage to the carrier sheet.

24A-24D illustrate the first alignment member 362 having first and second contact regions 377, 378 and the second alignment member 364 having a third contact region 379, other arrangements can be used. For example, the second alignment member 364 can include two contact areas, and the first alignment member 362 can include one contact area. Further, while the alignment members 362, 364 are illustrated as having irregularly shaped geometries for providing the first, second, and third contact regions 377, 378, and 379, other geometries may also be suitable for providing the first, second, and third contact regions. In other embodiments, the alignment members 362, 364 can provide more than three separate (e.g., discrete) contact areas for engaging the slide 243.

Referring back to fig. 24B, in the aligned position, the longitudinal axis 271a of the slide 243 can be substantially aligned (e.g., parallel) with the longitudinal axis 273a of the holding region 250 a. After aligning the slide 243, the alignment members 362, 364 can be disengaged from the slide 243 and returned to the open position by moving the blocks 365, 366 in a direction opposite to the direction of arrows 375, 376 (fig. 24A). Optionally, the staging device 210a can include a sensor 382 or other signal device for determining the presence of the slide 243 on the standby platform 240a and/or determining when the longitudinal axis 271a is substantially aligned with the longitudinal axis 273a (fig. 24B). For example, the standby platform 240a and/or the holding area 250a may include position sensors, pressure sensors, light sensors, etc. for determining the relative position of the slide 243 with respect to the holding area 250 a. Similar to the configuration and operation of the alignment device 242 (fig. 16-18), the alignment device 242a can be configured to align slides having different sizes and align them to desired locations on the standby platform 240 a.

After aligning the slide 243, the slide can be retrieved and transported to a specimen processing station (not shown). Fig. 25 and 26 illustrate a portion of a transport assembly 410 having a slide transfer head 412 ("transfer head 412"), the slide transfer head 412 configured to pick up an aligned slide 243 from the standby platform 240a while maintaining proper alignment. Referring to fig. 25, transfer head 412 includes a plurality of head alignment features 413 (e.g., 2 head alignment features) on a lower surface 415 of transfer head 412. The head alignment feature 413 can include, but is not limited to, a pin (e.g., an elongated rod), a protrusion, an opening (e.g., an opening defined by a bushing, an opening in a plate, etc.), and the like. In some embodiments, the head alignment features 413 can be in the form of alignment pins (e.g., first and second alignment pins) that can be inserted into corresponding alignment features 414 (shown individually as 414a and 414 b) on the staging device 210a (e.g., on the cantilevered plate 248 a), as shown in fig. 22 and 25. In other embodiments, the head alignment feature 413 is an opening and the corresponding alignment feature 414 is an upwardly projecting pin. In some embodiments, transfer head 412 can be a floating head (e.g., not contacting the floating head of staging device 210 a) to limit or prevent binding between head alignment features 413 and corresponding alignment features 414. In some embodiments, the transfer head 412 and/or staging device 210a can include position sensors (not shown) to ensure proper alignment of the head alignment features 413 relative to the corresponding alignment features 414.

The transfer head 412 can also include one or more capture features 416. The capture features 416 can include, but are not limited to, one or more suction devices (e.g., suction cups, pumps, vacuum pumps, etc.), mechanical grippers (e.g., grippers, clamps, tweezers, magnets, etc.), or other retention features that, for example, prevent the slide 243 from being dropped and/or transferred in a misaligned state. For example, transfer head 412 can include vacuum port 417 on lower surface 415. The vacuum source 418 can provide suction at the vacuum port 417 via the supply conduit 419 that can pick up the slide 243 from the staging device 210a and hold the slide during further transport. The vacuum can be reduced and/or eliminated to release the slide 243 after transfer. Sensors 405 (e.g., pressure sensors, air pressure sensors, light sensors, etc.) that detect the presence of a slide 243 held by the transfer head 412 can be disposed on the lower surface 415 and/or within the vacuum port 417, vacuum source 418, and/or supply conduit 419.

Fig. 25 shows the transfer head 412 in a non-engaged position over the staging device 210a during the alignment stage of slide transfer. The head alignment features 413 are shown aligned with corresponding alignment features 414 a. Fig. 26 shows the transfer head 412 lowered (e.g., by a drive mechanism, not shown) into an engaged position above the staging device 210 a. Head alignment features 413 (e.g., pins) are shown received within openings of corresponding alignment features 414 a. The vacuum port 417 is shown engaged with an upper surface 247 of the slide 243 (e.g., a label of the slide 243) such that when the vacuum source 418 is activated (e.g., by the controller 144 of fig. 1 and 2) and the over-travel inhibitor 254a associated with the standby platform 240a is disengaged (e.g., the vacuum provided by the stage vacuum source 281a is reduced and/or eliminated), the slide 243 can be picked up by the transfer head 412. The slide 243 can be removed from the staging device 210a when the transfer head 412 is raised to the non-engaging position above the staging device 210 a. As shown in fig. 26, the head alignment features 413 are aligned with the corresponding alignment features 414 so that the slide 243 can be maintained in an aligned position during slide pickup. After removing the slide 243 from the staging device 210a, the transfer head 414 can transport the slide 243 to a specimen processing station (not shown).

Fig. 27 is a block diagram illustrating a method 1000 for transferring a specimen slide using the specimen processing system 100 described above and with reference to fig. 19-26. Referring to fig. 19-27 together, the method 1000 can include moving the specimen slide 243 from the slide carrier 170 (fig. 14) to the standby platform 240a of the staging device 210a (block 1002). By engaging the ejector element with the slide 243, the slide 243 can be moved using the ejector 212 to push the slide onto the slide holding area 250a of the standby platform 240 a. The method 1000 can further include drawing a vacuum through the over travel inhibitor 254a to stop forward movement of the slide 243 on the slide holding region 250a (block 1004). The method 1000 can also include detecting the presence of a slide 243 on the holding area 250a (block 1006). In some embodiments, the presence of the slide 243 can be detected by the controller 144 by a change in the vacuum suction (vacuum suction) of the over travel inhibitor 254 a. For example, the sensor 403 (fig. 25 and 26) can be configured to detect changes in pressure within the vacuum port 290, fluid conduit 283, and/or vacuum source 281 (see fig. 16). In other embodiments, the presence of a slide on the standby platform 240a can be detected using other sensors 382 (e.g., pressure sensors, light sensors, motion sensors, etc.). For example, the standby platform 240a can include one more sensor 382 (e.g., a position sensor, a pressure sensor, a light sensor) for detecting the presence of the slide 243. The method 1000 can also include aligning the slide 243 from the unaligned position to the aligned position (block 1008). For example, the actuator can move the alignment members 362, 364 toward the slide 243 such that the first, second, and third contact regions 377, 378, and 379 engage the slide to move the slide to the aligned position. After alignment of slide 243, the actuator can move the alignment members 362, 364 back to the starting position and away from the aligned slide. The method 1000 can further include transporting the slide 243 from the standby platform 240a to, for example, a specimen processing station while maintaining the slide alignment (block 1010). For example, the transport assembly 410 with the transfer head 412 can be aligned with the standby platform 240a by alignment of the head alignment feature 413 on the transfer head 412 with the corresponding alignment feature 414 on the standby platform 240 a. The transfer head 412 can be configured to engage, pick up, and transport the slide 243 using the capture features 416. In one embodiment, the capture features 416 can use a vacuum provided by a vacuum source 418 through a vacuum port 417.

Fig. 28 and 29 show opposable dispenser 380 that includes opposable carrier holder 384 ("holder 384") and conveyor system 390. The transfer mechanism is capable of transporting the opposable carrier from the loading station 130 (fig. 1) to the holder 384. In some embodiments, including the illustrated embodiment, the holder 384 is configured to hold four magazines 391a, 391b, 391c, 391d (collectively "391") that each hold 30 opposable items to provide an on-board capacity of 120 opposable items. In other embodiments, the dispenser 380 can hold a greater or lesser number of cassettes or other types of opposable carriers.

The conveyor system 390 includes a carriage 393, a track 396, and an actuation mechanism 398. The actuation mechanism 398 can include an actuator (e.g., a piston assembly, a cylinder, etc.) that moves the vertical lift 404 to raise and/or lower the cartridge 391. The carriage 393 can carry the lowered opposable magazine to an unloading position at the end of the track 396. Fig. 28 and 29 show the empty cassette 394 in an unloaded position. The vertical lift 404 moves upward to retrieve the next cassette 391 and the carriage 393 moves the empty cassette 394 below the stack of cassettes 391. The carriage 393 is able to release the empty cartridge 394 such that the cartridge 394 drops the chute 397 into the storage bin 399 (shown in phantom).

Fig. 30 shows a transport assembly 420 and a specimen processing station in the form of a slide processing station in the form of a wetting module 430. The slides can be individually processed at the wetting module 430 to avoid carryover of liquids, excess waste (e.g., reagent waste), and/or reagent degradation to provide consistent processing. Wetting module 430 can stimulate the liquid using opposable element 470 to enhance process consistency, reduce process time, and allow processing with low concentrations of reagents. Relatively small volumes of reagents can be used to uniformly stain specimens. The specimen can be thoroughly washed in a relatively short period of time using a relatively small volume of wash solution. The wash cycle can be performed before, during and after the dyeing cycle. After processing the specimen, the transport assembly 420 can replace the used opposable 470 with a new opposable 457 and the used slide 243 with a new slide 458.

The transport assembly 420 can include, but is not limited to, a drive mechanism 434 (e.g., a rack drive mechanism, a belt drive mechanism, etc.) and a lift mechanism 440. The drive mechanism 434 is capable of moving the lift mechanism 440 horizontally, as indicated by arrows 450, 452. The lift mechanism 440 is capable of vertically moving an end effector in the form of a transfer head 454, 456, as indicated by arrows 462, 464. The delivery head can include, but is not limited to, one or more suction devices (e.g., suction cups, pumps, vacuum pumps, etc.), mechanical grippers (e.g., grippers, clamps, etc.), retention features (e.g., features that prevent the slide/dockable item from falling), and the like. For example, transfer head 454 can be a pick-up head (e.g., a rotatable or floating pick-up head) capable of picking up and holding opposable 457 by vacuum. The vacuum can be reduced (e.g., eliminated) to release the opposable 457. Additionally or alternatively, the mechanical gripper can hold the opposable 457.

Fig. 31 shows the delivery heads 454, 456 delivering opposable 457 and slide 458, respectively, to the wetting module 430. The transfer head 456 includes head alignment features 490, 492 that are receivable by the complementary alignment features 500, 502 (fig. 30) of the backup platform 240 and/or the alignment features 510, 512 (fig. 30) of the wetting module 430. The alignment features can include, but are not limited to, pins (e.g., elongated rods), protrusions, openings (e.g., openings defined by bushings in the plate, openings, etc.), and the like. In some embodiments, the alignment features 490, 492 are in the form of pins that can be inserted into the respective alignment features 510, 512 in the form of openings to align the slide 243 with the wetting module 430. The transfer head 456 can be a floating head to limit or prevent binding between the alignment features 490, 492 and the alignment features 510, 512, respectively. In other embodiments, the alignment features 490, 492 are openings and the alignment features 510, 512 are upwardly projecting pins.

After removing the treated slides 243, the transfer head 456 can transport the untreated slides 458 from the staging device to the wetting module 430. The alignment features 490, 492 can be positioned over the alignment features 510, 512, and the transfer head 456 can be lowered to insert the alignment features 490, 492 into the alignment features 510, 512, respectively, until the slide 458 rests on the wetting module 430. The transfer head 456 is capable of releasing the slide 458. After processing the specimen, the transfer head 456 can retrieve and load another slide into the wetting module 430. The slides can be held at the wetting module 430 to prevent damage to the slides in the event of a power outage or in other circumstances that can affect system performance.

After removing used opposable 470, transfer head 454 can deliver opposable 457 to opposable receiver 480. Once opposable 457 is positioned above wetting module 430, transfer head 454 can rotate opposable 457 from a substantially horizontal orientation (fig. 30) to a substantially vertical orientation (fig. 31). In some embodiments, opposable 457 oriented substantially horizontally defines an angle of less than 5 degrees from an imaginary horizontal plane and opposable oriented substantially vertically defines an angle of less than 5 degrees from an imaginary vertical plane. The vertically oriented opposable 457 can be loaded into the opposable receiver 480. The delivery head 454 can remove a used opposable and retrieve an unused opposable from an opposable carrier (e.g., the opposable carrier holder 384 of fig. 28 and 29), and can load the unused opposable into the opposable receiver 480.

Fig. 32 shows opposable actuator 525 including opposable receiver 480 and drive mechanism 530. Opposable receiver 480 can include clamp 536 and body 540. Clamp 536 includes a pair of clamps 542A, 542B that cooperate to hold mounting ends 950 of opposable 470. The opposable 470 includes a body 541 extending to the suction end 543. Body 541 is pivotally coupled to drive mechanism 530 by pivot 550. The drive mechanism 530 can include a linkage assembly 560 and a linear actuator assembly 562. The coupling assembly 560 includes a pivot 550 that allows rotation about one or more axes of rotation (e.g., two axes of rotation), and can include one or more ball bearings, pivots, hinges, or other features that provide the desired motion. The linear actuator assembly 562 can include an energizable drive 570 (e.g., stepper motor, drive motor, solenoid, etc.), a movable member 572 (e.g., lead screw, drive rod, etc.), and a track assembly 574 (e.g., carriage/track assembly, cage ball bearing linear track assembly, etc.).

Opposable receiver 480 is actuatable via linkage assembly 560 by linear actuator assembly 562. Linear actuator assembly 562 can be retracted and one or more fixed cams (e.g., cam 575 of fig. 33) can engage pins 576, 578 and drive opposable receiver 480 to the open configuration. In some embodiments, including the illustrated embodiment of fig. 32, opposable receiver 480 in the open configuration is capable of loosely retaining opposable 470. The opposable receiver 480 can be moved to the closed configuration by one or more biasing members (e.g., springs, pneumatic actuators, etc.). As the linear actuator assembly 562 extends, the pins 576, 578 are able to move upward and toward each other such that the biasing member closes the opposable receiver 480.

Opposable actuator 525 can also include, but is not limited to, one or more sensors that detect the presence of opposable 470, the position of opposable 470, one or more characteristics of treatment liquid engaged by opposable 470, and the like. These sensors can include, but are not limited to, contact sensors, electromechanical sensors, optical sensors, or chemical sensors that can be coupled to or incorporated into opposable receiver 480 or other suitable components. The number, location and configuration of the sensors can be selected to achieve the desired monitoring function.

Fig. 33 is an isometric view of a wetting module 430 holding a slide 243 in accordance with embodiments of the present technique. The wetting module 430 includes an opposable actuator 525, a slide holder platen 601, and a manifold assembly 606. Opposable actuator 525 in a rolling operating state can be extended or retracted to roll opposable 470 back and forth along slide 243. Movement of the swivel joint of the coupling assembly 560 (fig. 32), gravity, and/or liquid capillary forces can help maintain the desired movement of the opposable 470. In some embodiments, opposable actuator 525 can cause opposable 470 to continuously or periodically roll (e.g., longitudinally roll, laterally roll, or both), to agitate a volume of liquid, to move (e.g., translate, diffuse, narrow, etc.) a band of liquid (e.g., a meniscus layer of liquid), to control evaporation (e.g., mitigate evaporation), and/or to otherwise manage a treatment liquid.

The manifold assembly 606 includes a pair of sensors 620a, 620b (collectively "620") and one or more valves 630. The sensor 620 can detect a pressure of the working fluid and can send one or more signals indicative of the detected pressure. The fluid conduit 638 can fluidly couple the pressurized source 640 to the manifold 641. Fluid conduits 642, 644 fluidly couple the manifold 641 to the liquid removal device 655 and the slide holder platen 601. The liquid removal device 655 is capable of removing liquid between the opposable 470 and the slide 243 via the waste port 643. The conduit 644 can be used to draw a vacuum to hold the slide 243 on the slide holder platen 601.

Fig. 34A and 34B are isometric views of a slide holder platen 601 in accordance with embodiments of the present technology. The slide holder platen 601 of fig. 34A supports a slide 243. The slide holder platen 601 of fig. 34B is empty. The slide holder platen 601 can include a support element 650 and a mounting base 651. The support element 650 includes a raised chip receiving region 680 having a contact portion or contact surface 679 (fig. 34B). The port 683 (fig. 34B) is positioned to draw a vacuum to hold the slide 243 against the contact surface 679. The port 683 can be a suction cup or other feature configured to facilitate drawing a strong vacuum between the slide 243 against the contact surface 679.

The support element 650 includes an inner wall 681 positioned in the outer wall 652 of the mounting base 651. The inner wall 681 and the outer wall 652 form a heatable sidewall 682. In some embodiments, the sidewalls 682 can be positioned on both sides of the contact surface 679 and can output thermal energy to the ambient air to control the temperature of the slide 243, the processing fluid, and/or the specimen(s). In some embodiments, the sidewall 682 can also be positioned to laterally surround the entire slide 243. The mounting base 651 can be made of an insulating material (e.g., plastic, rubber, polymer, etc.) that can insulate the support element 650 from other components. In some embodiments, the mounting base 651 is made of a material having a thermal conductivity such that: which is substantially less than the thermal conductivity of the material of the support element 650. The mounting base 651 can surround and protect the support element 650 and includes a coupling region 657 to which the opposable actuator 525 can be coupled.

The support element 650 can be an uncoated element comprising one or more low thermal transfer materials having low thermal conductivity. The low heat transfer material can include, but is not limited to, steel, stainless steel, or other material having a thermal conductivity in the range of about 10W/(m × K) at 25 ℃ to about 25W/(m × K) at 25 ℃. In one embodiment, the low thermal transfer material comprises stainless steel having a thermal conductivity of 16W/(m × K) at 25 ℃. In some embodiments, support element 650 comprises primarily stainless steel by weight. In certain embodiments, at least a majority of the material of the support element 650 directly between the heating element 653 (fig. 35) and the slide 243 comprises stainless steel by weight. The stainless steel support element 650 can be corrosive resistant to the liquids used to process the specimens to provide a relatively long working life. In some embodiments, support element 650 comprises antimony (K = 18.5W/(m K) at 25 ℃) or chrome nickel steel (e.g., 18% Cr and 8% Ni by weight, and having a thermal conductivity of about 16.3W/(m K) at 25 ℃). In other embodiments, the support element 650 can include lead having a thermal conductivity of about 35W/(m × K) at 25 ℃ or other metals having similar thermal conductivities. In some embodiments, the support element 650 can be made of a material having a lower thermal conductivity than copper or brass. The mounting base 651 can be made of an insulating material having a thermal conductivity less than that of the support element 650. In this manner, the mounting base 651 can thermally insulate the support element 650.

Fig. 35 is a front, bottom, left side view of slide holder platen 601. Fig. 36 is a bottom view of slide holder platen 601. The slide holder platen 601 can include a heating element 653 that can convert electrical energy to thermal energy, and can include, but is not limited to, one or more traces (traces), leads, resistive elements (e.g., active elements that generate thermal energy), fuses, and the like. In some embodiments, the heating element 653 can be a resistive heater. Other types of heaters can also be used if needed or desired. In some embodiments, the heating element 653 can output thermal energy to the support element 650 to achieve a desired heat transfer pattern. Heat can be transferred non-uniformly to the slide 243 through the support element 650 to compensate for evaporative heat loss. The non-uniform heat transfer along the contact surface 679 may produce a non-uniform temperature profile along the contact surface 679. A substantially uniform temperature profile can be generated across the processing zone 671 (fig. 34A) of the slide 243. The processing zone 671 can be a staining region, a mounting region, or a region of the upper surface of the slide 243 or specimen-bearing surface 687 (fig. 34A) adapted to carry one or more specimens.

The heating element 653 of fig. 36 can include two elongate slide heating portions 660a, 660b (collectively 660) and two end heating portions 665a, 665b (collectively "665"). The elongated portion 660 delivers thermal energy to the longitudinally extending edge portion of the slide 243. The end-heated portion 665 delivers thermal energy to an end of the processing zone 671. The elongated portion 660 and the end heated portion 665 can be coupled together to form a multi-piece heating element 653. The elongated portion 660 and the end heating portion 665 can be made of materials having the same thermal conductivity or different thermal conductivities. Each section 660, 665 can be independently operated to output different amounts of thermal energy. In other embodiments, the heating element 653 can have a one-piece construction with a uniform thickness or a variable thickness. The one-piece heating element 653 can be made of one material.

The elongated portions 660 and the end heated portions 665 together define convective cooling features in the form of recesses 670. The recess 670 can help isolate heat in the support element 650 to help maintain thermal energy at the location where it is applied, and can also help reduce or limit the thermal mass of the slide holder platen 601. The recess 670 can be an opening having a substantially rectangular shape, as shown in fig. 36. However, the recesses 670 can have other shapes based on the desired heat distribution along the contact surface 679 of the support element 650.

Figure 37A is a cut-away isometric view of slide holder platen 601. The bearing element 650 includes a receiving region 680, a sidewall 682, and a passageway 684. The receiving region 680 keeps the slide 243 separate from fluids that can collect in the channel 684 during operation. The channel 684 is capable of collecting liquid that falls from the edges 813, 815 of the slide 243. In some embodiments, the slide 243 can extend outward from the receiving region 680 a sufficient distance (e.g., 0.5 mm, 0.75 mm, 1 mm, 2 mm, 4 mm, or 6 mm) to prevent capillary action of liquid between the slide 243 and the contact surface 679.

The slide holder platen 601 can be made in a multi-step manufacturing process. The support element 650 can be formed by a machining process, a stamping process, or the like. The support element 650 can be overmolded to form a mounting base 651, which can be made of an insulating material molded using an injection molding process, a compression molding process, or other suitable manufacturing process. Exemplary non-limiting insulating materials include, but are not limited to, plastics, polymers, ceramics, and the like. The support element 650 and the mounting base 651 can remain securely coupled together to inhibit or prevent liquid from traveling between the support element 650 and the mounting base 651. For example, the interface between support element 650 and mounting base 651 can form a fluid-tight seal with or without any sealant. However, sealants, adhesives, and/or fasteners can be used to securely couple the support element 650 to the mounting base 651. The illustrated support element 650 includes locking features 690, 692 to help minimize, limit, or substantially prevent movement of the support element 650 relative to the mounting base 651.

Figure 37B is a cross-sectional view of slide holder platen 601. Opposable 470 engages liquid 802, and the liquid 802 engages specimen 807. The sidewall 682 can extend vertically through the slide 243. The distance that the sidewall 682 extends vertically through the slide 243 can be selected to manage (e.g., limit, minimize, substantially prevent, etc.) airflow that can cause heat loss through convection (e.g., convection through ambient air), evaporation, and the like. For example, the slide holder platen 601 and opposable 470 can mitigate evaporation by maintaining the evaporation rate of the liquid 802 at or below about 7 microliters per minute, about 5 microliters per minute, about 3 microliters per minute, or other maximum evaporation rates. In some embodiments, the slide holder platen 601 and opposable 470 can maintain an evaporation rate of the liquid 802 in a range of about 7 microliters per minute to about 1 microliter per minute. Such an embodiment can mitigate evaporation losses. The sidewall 682 and opposable 470 help to substantially thermally isolate the specimen from the surrounding environment. In addition, the sidewall 682 can heat air proximate the specimen to help prevent the liquid 802 from being cooled by the surrounding air and inhibit or help prevent condensation.

The side portion 811 of the opposable 470 extends outwardly past the edge 813 of the slide 243 such that the side portion 811 is closer to the sidewall 682 than the edge 813 of the slide 243. Width W of gap 819_G1Can be less than the distance D from the side 811 to the edge 813 of the slide₁. Side portions 812 of opposable 470 extend outwardly past edge 815. Width W of gap 817_G2Can be less than the distance D from the side 812 to the slide edge 815₂. In some embodiments, the width W_G1Can be equal to or less than about 10%, about 25%, or about 50% of the distance between the left sidewall 682 and the rim 813. Similarly, width W_G2Can be equal to or less than about 10%, about 25%, or about 50% of the distance between the right sidewall 682 and the slide edge 815. Width W_G1、W_G2Can be small enough to inhibit or limit evaporative losses while allowing slight side-to-side movement of the item 470 to facilitate convenient handling. In some embodiments, the width W_G1、W_G2Equal to or less than about 1 mm, about 2 mm, about 4 mm, or other suitable width.

Fig. 38 is a top plan view of a wetting module 430. Fig. 39 is a cross-sectional view of a portion of the wetting module 430 taken along line 39-39 of fig. 38. Fig. 40 is a cross-sectional view of a portion of the wetting module 430 taken along line 40-40 of fig. 38. Referring to fig. 38 and 39, a sensor 694 is positioned to detect liquid in a reservoir 697. The sensor 694 can include a thermistor element 695 positioned near a bottom 696 of the reservoir 697. When a sufficient volume of liquid is collected to contact the thermistor element 695, the sensor 694 sends a signal to the controller 144 (FIG. 2). Detection of a threshold volume of liquid in reservoir 697 can indicate a fault in wetting module 430. Upon detecting a fault, wetting module 430 can be disabled until wetting module 430 can be, for example, inspected, cleaned, or otherwise serviced.

Referring to fig. 39 and 40, the wetting module 430 includes a convection system 700 including a flow generator 710, a conduit 711, and a flow path 712 (illustrated in phantom) defined by a passageway 713 through the conduit 711. The flow generator 710 can include, but is not limited to, one or more fans, blowers, or other suitable components capable of generating a sufficient flow of convective fluid (e.g., air, refrigerant, etc.) along the flow path 712 to cool the back of the support element 650, the slide 243, and/or items (e.g., specimens, reagents, etc.) carried on the slide 243.

The flow generator 710 can deliver convective fluid toward an end 730 of the support element 650 that is below a first end 732 of the slide 243. The convective fluid can travel vertically through the tapered section 720, which tapered section 720 can accelerate the flow of the convective fluid. The accelerated stream is directed horizontally and flows under the slide platen 601. The convective fluid can directly contact the support element 650 to facilitate and accelerate cooling of the slide 243. For example, a convective fluid can flow into and along the recess 670 to absorb thermal energy from the support element 650. The support elements 650 absorb thermal energy from the slide 243 to cool the upper surface 687 and ultimately the liquid, specimen, or any other item or substance on the upper surface 687. The warmed fluid flows through the recess 670 and continues to travel under the end 750 of the support element 650 below the label end 752 of the slide 243. The air flows down through the outlet 760 to the ambient environment.

The convection system 700 can be used to rapidly cool the slide 243. For example, the convection system 700 can facilitate cooling the liquid and/or specimen at a rate equal to or greater than about 2.5 ℃/sec. In one embodiment, the temperature of the specimen can be at about 95 ℃ and can be cooled to a temperature equal to or less than about 30 ℃ in about four minutes or less. Other cooling rates can be achieved by increasing or decreasing the flow rate of the convective fluid, the temperature of the convective fluid, etc. During the heating cycle, the convection system 700 can be OFF, if desired.

Fig. 41 is a cross-sectional view of a portion of the slide holder platen 601 taken along line 41-41 of fig. 38. The temperature of the liquid 802 can be maintained within a target temperature range selected based on the characteristics of the liquid 802, the characteristics of the specimen (e.g., thickness of the specimen, composition of the specimen, etc.), and the process to be performed. Because the area of the liquid 802 closest to the edge of the slide 243 evaporates more than the central area of the liquid 802, the periphery of the slide 243 and the periphery of the liquid 802 tend to be at a lower temperature without compensation. The evaporative heat loss for high temperature processes (e.g., antigen retrieval) may be greater than the evaporative loss for low temperature processes (e.g., rinsing). Because significant temperature variations along the specimen 807 and/or the liquid 802 can result in process variations, the wetting module 430 can maintain a desired temperature profile of the slide 243 by compensating for evaporative heat losses, including evaporative heat losses during high and low temperatures. The wetting module 430 is capable of generating a substantially uniform temperature profile along the surface 687 to substantially uniformly heat the ribbon of liquid 802 and/or the specimen 807. A uniform temperature profile can be maintained independent of changes in the ambient environment to consistently process the entire specimen 807.

Figure 41A is a plot of position along the width of the receiving region 680 versus thermal energy conducted to the slide 243. Fig. 41B is a graph of the location along the width of the receiving area 680 versus the temperature of the contact surface 679 of the support element 650. Fig. 41C is a view of the location along the upper surface 687 of the slide 243. A comparison of fig. 41B and 41C shows that the temperature profile along the contact surface 679 of the support element 650 differs from the temperature profile along the upper surface 687 of the slide 243.

Referring to fig. 41A, the heating element 653 can deliver thermal energy non-uniformly by conduction to the slide 243. The heat is still concentrated at the perimeter of the dye area where the evaporation heat loss is relatively high. Because thermal energy is not directly transferred by conduction to the portion of the support element 650 above the recess 670, a non-uniform temperature profile is generated along the contact surface 679 of the support element 650 and can compensate for non-uniform heat loss associated with evaporation of the liquid 802. The compensation can result in a substantially uniform temperature profile along the upper slide surface 687. As shown in fig. 41C, the temperature along the upper blade surface 687 can be kept within the target temperature range (indicated by two horizontal dashed lines). In embodiments for antigen retrieval, the substantially uniform temperature profile can have a temperature variation equal to or less than 5% of the desired temperature and can pass over a substantial portion of the upper carrier sheet surface 687. The upper carrier sheet surface 687 can be maintained at an average or target temperature of, for example, about 95 c and within a range of about 90.25 c and about 99.75 c. In some embodiments, the heater element 653 produces a temperature change of less than about 4% across a majority of the upper blade surface 687. In other embodiments, there can be less than 5% temperature variation across a majority of the upper blade surface 687. The upper carrier sheet surface 687 can be maintained at an average temperature of, for example, about 95 c and within a range of about 92.63 c and about 97.38 c. In some embodiments, the allowable temperature change can be input by a user.

FIG. 42 is a top view of a heating zone in accordance with embodiments of the present technique. The high heating zone 820 surrounds the intermediate heating zone 824. The intermediate heating zone 824 surrounds the lower heating zone 822. Heat from the heating element 653 travels primarily upward to define the high heating zone 820. The high heating zone 820 can be located below the perimeter of the stained area of the slide 243. The low heating zone 822 can generally correspond to the recess 670 and a central processing region (e.g., a staining region) where the specimen or specimens are generally located. The temperature of the heating zones 820, 822, 824 can be approximately inversely proportional to the evaporation rate of the slide directly above the heating zones. For example, the low heating zone 822 can be positioned substantially below the middle of the belt where substantially no evaporative loss of liquid 802 occurs. The high heating zone 820 is positioned substantially below the periphery of the band of liquid 802 that experiences relatively high evaporative losses.

Figure 43 is a flow diagram illustrating a method 900 for heating a slide, in accordance with embodiments of the present technology. At 901, the specimen-bearing slide 243 (fig. 34A) can be positioned on a contact surface 679 (fig. 34B) of the support element 650. The slide 243 can be preheated by the slide holder platen 601. The liquid can be delivered onto the heated slide 243. Alternatively, the slide holder platen 601 can heat the slide 243 after delivering the liquid.

At 902, the opposable 470 is used to manipulate the liquid and can mitigate and control evaporation, which in turn can affect temperature, concentration, and capillary volume. In some embodiments, the liquid is allowed to evaporate, resulting in heat loss and, in some embodiments, a change in the concentration of the liquid 802. The dispenser can deliver supplemental liquid at a desired time to maintain the volume of liquid in a desired range, maintain a desired concentration of liquid, and the like. If the current liquid volume is below the target equilibrium volume, the controller can instruct the dispenser to deliver liquid until the current liquid volume reaches the equilibrium volume. If the current liquid volume is above the target equilibrium volume, the controller can instruct the dispenser to stop delivering liquid until the current liquid volume reaches the equilibrium volume. Once the liquid reaches the target equilibrium volume, the controller can instruct the dispenser to provide make-up fluid to the liquid at a desired rate (e.g., a fixed rate or a variable rate) in order to maintain the liquid at the equilibrium volume. The delivery rate can be selected based on the evaporation rate of the liquid.

At 903, the contact surface 679 can have a non-uniform temperature profile such that the upper surface 687 of the slide 243 has a more uniform temperature profile than the non-uniform profile of the contact surface 679. Substantially the entire mounting area of the slide 243 can have a substantially uniform curve. This ensures that any portion of the specimen contacting the mounting surface is maintained at a substantially uniform temperature for consistent processing. The specimen can be processed consistently even if it is moved slightly along the mounting surface.

At 904, heat loss associated with evaporation of the liquid 802 can be compensated for by creating a non-uniform temperature profile along the contact surface 679. The support element 650 and the heated sidewall 682 can be used to control the temperature of the slide 243.

Repeated manipulation of the fluid on the staining surface results in fluid mixing between different regions within the body of fluid in contact with the slide surface, both in terms of mass and thermal energy mixing. Thus, temperature uniformity control over the surface of the slide is affected by the interaction of: 1) a conductive heating element beneath the slide; 2) thermal mixing caused by fluid manipulation; and 3) evaporative heat loss relative to the ambient environment. Fluid manipulation is governed by factors such as manipulation speed and distance relative to a specified volume. Therefore, the thermal profile of the conductive elements under the slide must be properly designed for optimal temperature uniformity across the slide relative to fluid handling factors.

Fig. 44 shows the slide holder platen 601, the dispenser assembly 633, and the controller 144 of the evaporation-lightened specimen processing station. The dispenser assembly 633 includes a fluid source 621 fluidly coupled to the dispenser 622 via a fluid conduit 623. The fluid source 621 can include, but is not limited to, one or more containers (e.g., a container taken from the docking station 124 of fig. 1, a container taken from the docking station 142 of fig. 1, etc.), a reservoir or other suitable fluid source (e.g., a bulk reagent reservoir), and can include one or more valves, pumps, etc. The dispenser 622 is capable of outputting liquid via the array of conduits 625. In some embodiments, including the illustrated embodiment of fig. 44, the distributor 622 includes eight conduits 625, although any number of conduits can be used. Further, the dispenser assembly 633 can include more than one dispenser depending on the design of the slide holder platen 601. Additionally or alternatively, the dispensers 160, 162 of fig. 2 can deliver liquid onto the slides and can be fluidly coupled to the fluid source 621 or another fluid source. The opposable 470 can be positioned to allow one or both of the dispensers 160, 162 to deliver liquid onto the slide. In some embodiments, the dispenser 622 delivers bulk liquid from the container at the docking station 142 and the dispensers 160, 162 deliver liquid from the container at the docking station 140.

The controller 144 can control the array of specimen processing stations to maintain the volume of processing liquid within the equilibrium volume range. If the volume of the liquid is above the equilibrium volume range, the liquid can evaporate at a relatively high rate and the concentration of the liquid can be significantly altered. If the volume of liquid is below the equilibrium volume range, the volume of liquid may not be sufficient to adequately process the specimen. Furthermore, insufficient liquid volume can lead to an undesirably low amount of liquid agitation during processing. The equilibrium volume range can be selected based on the composition of the liquid, the desired processing temperature, or the desired agitation of the liquid 802. The equilibrium volume of the liquid 802 can correspond to a volume of fluid (at a particular temperature or temperature range) that provides complete coverage of the specimen while keeping evaporation losses below a target level. The dispenser 622 can function as a replenishment device that periodically replenishes liquid at a fixed rate (e.g., a rate based on the evaporation rate) to maintain the volume of liquid within an equilibrium volume range, replenish depleted reagent, and the like.

Using the target process temperature or target process temperature range and the total evaporation rate, the controller 144 can determine a target range for the equilibrium volume. In some embodiments, the controller 144 can receive the total evaporation rate information from the memory 629 and/or the input device 628. The input device 628 can include a data server or other similar device that can provide information from a database upon request or periodically. The total evaporation rate information can be obtained from empirical studies and stored in a database. In other embodiments, the input device 628 can be a reader that obtains information (e.g., target processing temperature range, replenishment rate, etc.) from the label of the slide.

The controller 144 can receive information (e.g., lookup tables, temperature set points, duty cycles, power settings, environmental information such as ambient temperature and/or humidity, processing protocols, etc.) from the memory 629. The input device 628 can be a manual input device (e.g., keyboard, touch screen, etc.) or an automated input device (e.g., computer, data storage device, server, network, etc.) that can provide information automatically upon request from the controller 144. The memory 629 can store different instructions for different processes. One stored sequence of program instructions can be used to contact the specimen 807 with the wash solution, and another sequence of program instructions can be used to apply a reagent (e.g., a stain) to the specimen. The controller 144 can include a programmable processor 631 that executes sequences of program instructions to sequentially process specimens with wash solutions and reagents. The slide holder platen 601 can heat the slide to a first target temperature when executing a first sequence of program instructions and can cool the slide to a second target temperature when executing a second sequence of program instructions. Any number of sequences of program instructions can be executed to perform the various stages of the scheme.

The controller 144 can also be programmed to control the wetting module 430 so that the dispenser 622 delivers the replenishment liquid onto the slide. The rate of fluid delivery can be based on, for example, processing information (e.g., protocol, agitation information, one or more processing times, etc.), total evaporation rate information (e.g., evaporation rate under particular conditions, actual evaporation rate for a particular type of liquid, etc.), and the like. The current liquid volume can be determined based on the initial liquid volume on the slide and the stored evaporation rate or rates. The stored evaporation rate can be input into the system 100 or determined by the system 100. The controller 144 can calculate the equilibrium volume in advance (e.g., a test run), and the system 100 can use the determined equilibrium volume as an initial volume for the same type of liquid. The controller 144 can then instruct the dispenser 622 to provide the supplemental liquid at a rate (e.g., a rate determined by a test run). In some embodiments, the rolling speed can be about 100 mm/s to provide a substantially uniform temperature profile. For example, a rolling speed of 100 millimeters per second can provide a temperature range on the slide of about 4.2 ℃, while a rolling speed of 65 millimeters per second provides a temperature range of about 6.2 ℃. The rolling direction, rolling speed and rolling frequency can be adjusted depending on the type of liquid and the desired temperature profile. The rolling speed can have a direct effect on the overall evaporation rate. A faster rolling speed can result in a higher evaporation rate. This can be a factor to consider when gathering demonstrated total evaporation volume information to generate a protocol.

The power supply 627 of the controller 144 can be electrically coupled to a heating element (e.g., heating element 653 of fig. 37A and 37B). The power source 627 can be one or more batteries, fuel cells, or the like. The power supply 627 can also deliver electrical energy to other components of the system. In other embodiments, the power supply 627 can be an AC power supply.

Fig. 45 and 46 are perspective and top views, respectively, of another embodiment of a slide holder platen 701 shown with a slide 243 and configured in accordance with the present techniques. Fig. 47 is a perspective view of slide holder platen 701 without slide 243. Referring to fig. 45-47, slide holder platen 701 is substantially the same as slide holder platen 601 discussed above in connection with fig. 34A-44, except as explained in detail below. Slide holder platen 701 can include support element 703, sealing member 709, and vacuum port 721. The support element 703 includes a raised slide receiving region 707, and the sealing member 709 is configured to engage a bottom surface of the slide 243 when the slide is placed on the slide receiving region 707. The sealing member 709 can be positioned around the vacuum port 721 such that when the slide 243 engages the sealing member 709, a vacuum is drawn via the vacuum port 721 to pull the slide 243 against the sealing member 709 to maintain a seal (e.g., an airtight seal) and prevent or limit unwanted movement (e.g., rotational and/or translational movement as indicated by arrows 801a-b and 799a-b, respectively, in fig. 46) of the slide 243 relative to the slide receiving region 707.

Referring now to fig. 47, the slide receiving area 707 can have a first portion 733 and a second portion 735 disposed within an opening 745 of the first portion 733. The vacuum port 721 can be disposed at a top surface 735a of the second portion 735 at a substantially central location. The vacuum port 721 can be fluidly coupled to a vacuum source 717 via one or more fluid conduits 719 (e.g., internal fluid conduits, external fluid conduits, etc.). For example, one or more fluid conduits 719 can extend through the second portion 735 from the opening 705 at the top surface 735a to the vacuum source 717. Vacuum source 717 can include, but is not limited to, one or more pressurization devices, pumps, or other types of devices capable of drawing a vacuum through opening 705. As shown in fig. 46, when the slide 243 is positioned on the slide receiving region 707, the specimen-bearing portion 729 of the slide 243 is generally aligned with the first portion 733 and the label-bearing portion 723 of the slide 243 is generally aligned with the second portion 735. In this manner, the vacuum created by the vacuum port 721 can be confined to the label bearing portion 723 of the slide 243 to avoid disrupting the thermal treatment of the specimen bearing portion 729.

The second portion 735 and the opening 745 can individually have a non-circular shape (when viewed from above). As used herein, "non-circular" refers to any shape other than a true circle (i.e., a shape having a substantially constant radius at each point around its perimeter). For example, in some embodiments, the second portion 735 and/or the opening 745 can have a rectangular shape with rounded corners. In other embodiments, the second portion 735 and/or the opening 745 can have any non-circular shape, size, and/or configuration, such as a rounded polygonal shape, a polygonal shape, an oval, an ellipse, and the like. In some embodiments, including the illustrated embodiment, the second portion 735 and the opening 745 can have substantially the same non-circular shape, and in some embodiments, the second portion 735 and the opening 745 can have different non-circular shapes.

Fig. 48 is a partially exploded view of slide holder platen 701, and fig. 49 is a cross-sectional side view of a portion of platen 701 in fig. 48. Referring to fig. 48 and 49 together, the first portion 733 and the second portion 735 of the chip receiving region 707 are separated by a groove 737 that receives the sealing member 709. The groove 737 defines an opening 745 and can have an outer side wall 739 defined by the first portion 733, an inner side wall 741 defined by the second portion 735, and a floor portion 743 between the side walls 739, 741. Referring now to fig. 49, the height 775 of the outer sidewall 739/first portion 733 can be greater than the height 773 of the inner sidewall 741/second portion 735. As described in more detail below with reference to fig. 54, when the slide 243 is positioned on the slide receiving area 707, the backside surface of the slide contacts the top surface or contact surface 733a of the first portion 733 and is separated from the top surface 735a of the second portion 735 by a distance 781. As such, the difference in elevation between the first portion and the second portion creates a vacuum cavity 757 (fig. 54) defined at least partially by the top surface 735a of the second portion 735 surrounding the vacuum port 721.

Fig. 50 and 52A are perspective and top views, respectively, of the sealing member 709, and fig. 51 is a cross-sectional end view of the sealing member 709 taken along line 51-51 of fig. 50. The sealing member 709 can be in the form of a non-circular flexible gasket having a body 747 and a lip 749 extending radially outward from the body 747. The sealing member 709 is movable between an uncompressed configuration 709UC for contacting a slide moving toward the slide receiving area 707 and a compressed configuration 709C (shown in phantom) for maintaining a hermetic seal. The body 747 can have an inner surface 761 configured to contact an inner sidewall 741 of the groove and an outer surface 767 configured to contact an outer sidewall 739 of the groove 737. The lip 749 includes a top surface 763 configured to engage the backside of a microscope slide when the slide is placed on the slide receiving region 707. The lip 749 can extend radially outward from the body 747 a distance less than the outer surface 767 of the body 747. As such, lip 749 does not necessarily contact outer sidewall 739 when sealing member 709 is positioned within groove 737.

As shown in fig. 52A, the sealing member 709 (or body 747) can have a non-circular shape when viewed from above (or along an axis substantially perpendicular to a top surface 763 of the sealing member 709). For example, in some embodiments, the body 747 can have a rectangular shape with rounded corners (e.g., fig. 52A). In other embodiments, the body 747 can have any non-circular shape, size, and/or configuration, such as a rounded polygonal shape, a polygonal shape (e.g., square (fig. 52B), triangular (fig. 52C), etc.), a "petal" configuration (e.g., fig. 52D), etc. The sealing member 709 can be made, in whole or in part, of rubber, Polytetrafluoroethylene (PTFE), silicone, nitrile, vinyl, neoprene, and/or other compressible or flexible materials capable of maintaining a desired seal.

Fig. 53 is a cross-sectional side view of the platen 701 when the slide 243 is positioned on the slide receiving region 707, but before the back side 243a of the slide 243 is in contact with the sealing member 709 in an uncompressed state. As shown in fig. 53, at least a portion of the body 747 is in contact with the inner side wall 741, the outer side wall 739, and the bottom plate portion 743 of the groove 737. The lip 749 is spaced from an outer side wall 739 of the groove 737 and extends upwardly from the groove 737 beyond the top surface 733a of the first portion 733. The lip 749 can also extend upwardly from the groove 737 beyond the horizontal plane (imaginary plane) defined by the top surface 733 a. For example, the lip 749 can extend a distance 753 from the top surface 733 a. As such, the lip 749 is configured to engage the back-side surface 243a of the slide 243 before the back-side surface 243a of the slide 243 contacts the top surface 733a of the first portion 733. In this manner, the sealing member 709 absorbs contact forces associated with placing the slide 243 on the slide receiving region 707 to facilitate transitioning of the slide 243 onto the slide receiving region 707.

Fig. 54 is a cross-sectional side view of the platen 701 after a slide 243 has been positioned on the slide receiving region 707 (e.g., the sealing member 709 is in a compressed state), and fig. 55 is an enlarged view of a portion of fig. 54. As shown in fig. 54, the back side surface 243a of the slide 243 contacts the lip 749 of the sealing member 709 and the top surface 733a of the first portion 733. Because of the height difference between the first portion 733 and the second portion 735, the backside surface 243a of the slide 243 is separated from the top surface 735a of the second portion 735 by a distance 781 (see fig. 55). As such, the pressurization port 721 is positioned below and spaced apart from the back side 243a of the slide 243 such that the top surface 735a of the second portion 735 and the back side surface 243a of the slide 243 at least partially define the vacuum chamber 757. For example, when the vacuum source is activated, fluid and/or air between the back side 243a of the slide 243, a portion of the sealing member 709 (e.g., the lip 749 and/or the outer surface 761 of the body 747), the inner sidewall 741, and/or the top surface 735a of the second portion 735 is drawn through the vacuum port 721 (as indicated by arrow 755). As a result, the slide 243 is pulled against the sealing member 709, thereby forming a seal. The seal fixes the positioning of the slide 243 relative to the support element 703 and substantially eliminates unwanted rotation and/or translation of the slide 243.

Lip 749 can be movable between an uncompressed configuration and a compressed configuration without contacting outer sidewall 739 of groove 737. As best shown in fig. 55, a gap 771 is maintained between the lip 749 of the seal member and the outer side wall 739 of the groove 737 even in the compressed configuration. For example, the lip 749 can be configured to deflect primarily in a direction perpendicular to the back-side surface 243a of the slide 243. The lip 749 can be sufficiently rigid to prevent any rotation of the slide 243 about a vertical axis. As such, the slide 243 can be rotationally fixed relative to the support surface. Although the lip 749 can be spaced from the outer sidewall 739 (in a compressed state), the lip 749 is configured to physically contact one or more sidewalls of the groove 737 to inhibit movement of the slide 243 relative to the support element 703. For example, as shown in fig. 56, a lip 749 or other portion of the sealing member 709 can be configured to physically contact an outer sidewall 739 of the groove 737 when the slide 243 is rotated about its vertical axis (e.g., at least about 2 degrees). Because of the non-circular shape of both the sealing member 709 and the opening 745 in the first portion 733, the outer sidewall 747 of the groove 737 limits rotation of the sealing member 709 (e.g., by applying a contact force CF), and thus, limits rotation of the slide 743.

The slide holder platen 701 can include additional features. For example, the slide holder platen 701 can include one or more sensors 759 (fig. 54) that detect the presence of a slide 243 to be detected and/or activate the vacuum source 717. In some embodiments, the slide holder platen 701 can include one or more sensors to monitor the pressure generated within the vacuum chamber 757. In a particular embodiment, the slide holder platen 701 can be in communication with a controller that can control the timing and/or magnitude of the vacuum source 717.

FIG. 57 is a graph of equilibrium volume of treatment liquid versus total evaporation rate, in accordance with embodiments of the present technique. The x-axis represents the equilibrium volume (EV in μ L) and the y-axis represents the total evaporation rate (TER in μ L/s). The line T1 and the line T2 represent the relationship between TER and EV at temperature T1 and temperature T2, respectively. In the illustrated embodiment, T1 is higher than T2. The controller 144 can receive the total evaporation rate information from the memory 629, the input device 628, or the like. The total evaporation rate information can be measured and stored in the memory 629. The total evaporation rate information can include evaporation rates for liquids at different concentrations. After the controller 144 receives the predetermined temperature (e.g., T1) and the total evaporation rate information (e.g., "a" μ L/s), the controller 144 can determine an EV value (e.g., "B" μ L) of the liquid based on the graph of fig. 57. Equation 1 corresponds to the relationship described in fig. 57. The slopes of line T1 and line T2 represent the temperature dependent evaporation constant (K) as follows.

TER = kx EV formula 1.

Once the equilibrium volume of liquid is determined, the controller 144 can compare it to the estimated volume of the slide and can instruct the dispenser 622 to supply supplemental fluid if needed. If the current liquid volume is below the target equilibrium volume, the controller 144 can instruct the dispenser 622 to provide more make-up liquid.

Figure 58 is a graph of time versus slide coverage in accordance with an embodiment of the disclosed technology. FIGS. 59A-63B illustrate one way to achieve the coverage depicted in FIG. 58 by: by moving the liquid 802 along the entire dye area 671 (excluding the label 907 and a portion of the rim, if desired) to provide full coverage by moving alternately between the opposite ends 732, 735 of the mounting area 671. Full coverage can help minimize, limit, or substantially prevent problems associated with under-wetting and over-wetting. In the event of insufficient wetting, the liquid 802 contacts less than the entire stained area 671, so that the specimen 807 may be at risk of not being contacted and thus not being processed/stained. In the event of over-wetting, the liquid 802 contacts more than the entire stained area 671 and may tend to drain from the slide 243. In subsequent processes, the liquid 802 may be at risk of ineffective liquid removal, resulting in reagent residue and associated degradation of stain quality. If the liquid 802 is a stain, the entire specimen 807 is contacted for consistent (e.g., uniform) staining. If the liquid 802 is a cleaning liquid, complete coverage ensures that the entire specimen 807 is thoroughly cleaned, particularly after reagent processing. The different stages of the method are discussed in detail below.

Fig. 59A and 59B are side and top views of the ribbon of liquid 802 held between opposable 810 and mounting region end 732 by an opposable actuator (not shown) at time 0 in fig. 58. A band of liquid 802 (e.g., meniscus layer, film, etc.) can be formed for the holder 810 and slide 243. The band of liquid 802 of fig. 59B is shown in dashed lines. The gap 930 (e.g., capillary gap) can have a minimum holding capacity of about 125 milliliters to about 200 milliliters. Other minimum and maximum containment capacities are possible if needed or desired. The minimum containment volume can be the minimum volume of liquid that can be contained in the gap 930 and effectively applied to the specimen 807, which specimen 807 can be located anywhere on the staining area 671. The maximum containment volume is the maximum volume of liquid that can be contained in the gap 930 without overfilling. The gap of varying height 930 is able to accommodate a greater range of liquid volumes than a gap of uniform height because the narrowed region of the gap 930 is able to accommodate a smaller liquid volume.

Opposable 810 is rolled along slide 243 to displace the band of liquid 802 in the direction of longitudinal axis 951 of slide 243 (indicated by arrow 961). In fig. 60A and 60B, the band of liquid 802 has been spread by moving one side 958 of the band of liquid 802 in the direction of the longitudinal axis 951 (corresponding to 0.25 seconds in fig. 58). A side 956 of the band of liquid 802 can remain at the edge 960 of the slide 243. In some embodiments, the band of liquid 802 can be from a narrowed width W_N1(FIG. 59B) diffusion to the diffused width W_S. Width W_N1、W_SCan be substantially parallel to a longitudinal axis 951 of the slide 243, and the length L of the band of liquid 802 can be substantially perpendicular to the longitudinal axis 951.

Fig. 61A and 61B show the band of liquid 802 after the band of liquid 802 has moved along the slide 243, which corresponds to 0.5 seconds in fig. 58. The band of liquid 802 is displaced using capillary action. Capillary action can include, but is not limited to, movement of the band of liquid 802 due to the phenomena: due to stickingThe phenomenon of liquid creeping (peel) spontaneously across the gap 930 due to adhesion, adhesion and/or surface tension. In some embodiments, the width W_SCan be substantially maintained while displacing the band of liquid 802. In other embodiments, the width W is while moving the band of liquid 802_SCan increase or decrease by less than 5%. In some embodiments, opposable 810 can have a non-uniform curvature or configuration to have a variable width W as the belt moves through the slide_S。

Fig. 62A and 62B show a band of liquid 802 at end 735, which corresponds to 0.75 seconds in fig. 58. One side 958 of the strip of liquid 802 can be drawn between an end 952 of opposable 810 and an end 735 of the mounting area 671. The label 907 can help attract the liquid 802. For example, label 907 can be made, in whole or in part, of a hydrophobic material. When opposable 810 is moved to the over-rolled position of FIG. 63A, width Ws of band of liquid 802 can be reduced to narrowed width W_N2This corresponds to 1 second in fig. 58. The width of the band of liquid 802 can be reduced while drawing substantially all of the liquid 802 at the end 970 of the gap 930. For example, at least 90% of the volume of liquid 802 can remain attracted. In some embodiments, at least 95% of the volume of liquid 802 can remain attracted. In still other embodiments, substantially all of the liquid 802 can remain attracted while reducing the width of the band of liquid 802.

Compressed width W_N2Can be substantially less than the width Ws such that the entire band of constricted liquid 802 is spaced from the specimen 807. In some embodiments, the narrowed width W_N2Can be equal to or less than about 50%, about 25%, or about 10% of the width Ws. Such embodiments may be particularly well suited for processing slides carrying one or more specimens. The narrowed band coverage exposes a relatively large area of the dyed area 671 while preventing capillary action or leakage of the liquid. In some embodiments, the width W_N2Can be equal to or less than about 40%, about 30%, or about 20% of the width Ws. Width W_N1Can be substantially equal to the width W_N2. Advantageously, the opposable actuator 525 can be operated to increase or decrease to provide a variable constriction of the band of liquid 802.

Opposable 810 of fig. 63A and 63B can be rolled back on slide 243 to move the band of liquid 802 to the position shown in fig. 59A. The opposable 810 can be rolled back and forth any number of times at a variable or constant rate to move the band of liquid 802 back and forth across the slide 243. If the liquid 802 is a wash liquid, the wash liquid can be passed quickly back and forth over the specimen 807 to provide thorough washing. If the liquid 802 is a stain, a band of the liquid 802 can be traversed over the specimen 807 to span the entire width W of the specimen 807_specProviding uniform staining (measured in a direction parallel to the longitudinal axis 951 of the slide 243). One or more wash cycles can be performed between dyeing cycles. Mixing on the slide can also be performed if needed or desired.

The treatment protocol may require different rolling speeds and different liquid volumes to meet various treatment criteria (e.g., chemical requirements, uptake requirements, solubility limits, viscosity, etc.). If the specimen 807 is a paraffin-embedded specimen, a relatively small volume of dewaxing solution (e.g., 12 microliters of xylene) can be delivered into the gap 930. The opposable 810 can be rolled (e.g., rolled along an imaginary plane spaced from the upper surface of the slide 243, rolled along the upper surface, rolled sideways, rolled longitudinally, etc.) or otherwise manipulated (e.g., rotated, translated, or both) to apply the liquid 802. After dewaxing, a relatively large volume of reagent can be delivered into the gap 930. For example, a volume of about 125 microliters to about 180 microliters of stain can be delivered into the gap 930. The stain is delivered to the specimen 807 and then subsequently removed.

The methods shown in fig. 59A-63B can be used to perform assay steps (e.g., assays for antibodies and chromogens). The determining step can be performed at a relatively low temperature. The slide holder platen 601 can maintain the specimen and/or processing liquid at a temperature in a range of about 35 ℃ to about 40 ℃. In one embodiment, the liquid and/or specimen is maintained at a temperature of about 37 ℃. A dispenser (e.g., dispenser 622 of fig. 44) is capable of delivering a supplemental liquid to maintain a target volume of about 30 microliters to about 350 microliters. In some versions, the dispenser delivers the supplemental liquid at a rate of about 4 microliters to about 5.1 microliters per minute to about 5.6 microliters per minute. In such embodiments, the volume of the liquid (e.g., liquid 802 of fig. 59A) can be maintained in a range of about 90 microliters to about 175 microliters over a period of about 15 minutes based on a relative humidity of about 10% -90%, an ambient temperature of about 15 ℃ to about 32 ℃, with an average slide temperature tolerance of about ± 1 ℃ and an opposable rolling speed of about 25 millimeters to about 60 millimeters per second. The evaporation rate may be approximately proportional to the rolling speed. If the scroll speed is about 20 millimeters per second, a replenishment rate of about 3.8 microliters per minute to about 4.2 microliters per minute can maintain a volume of about 115 microliters to about 200 microliters. If the scroll speed is about 40 millimeters per second, a replenishment rate of about 5.1 microliters per minute to about 5.6 microliters per minute can maintain a volume of about 115 microliters to about 200 microliters of the liquid 802. At high roll speeds of about 90 millimeters per second, the replenishment rate can be about 7.6 microliters per minute to about 8.4 microliters per minute to maintain a volume of about 110 microliters to about 200 microliters. Higher velocities are also possible, but depend on the gap height, opposable radius and fluid properties. Humidity and ambient temperature can affect the evaporation rate at low temperatures, but may not have a significant effect at elevated temperatures, e.g., at temperatures greater than 72 ℃.

For targeted repair, the roll speed can be about 100 millimeters per second, and the replenishment rate can be 72 microliters per minute. For antigen retrieval, the scroll speed can be about 180 millimeters per second, and the replenishment rate can be about 105 microliters per minute. Other replenishment rates can be selected based on process conditions.

As used herein, the term "opposable element" is a broad term and refers to, but is not limited to, a surface, tile (tile), strip, or another structure capable of manipulating one or more substances to process a specimen on a slide as described herein. The components of system 100 (fig. 1) use a wide range of different types of opposable elements. In some embodiments, the opposable element can include one or more spacers, gap elements, or other features for positioning the opposable element relative to the slide. In other embodiments, the opposable element can have a smooth surface (e.g., a non-planar fluid-handling surface) that is substantially free of spacers, gap elements, or the like, and can have a single-layer or multi-layer construction. The smooth surface can be rolled or otherwise advanced along the slide. As discussed above, the opposable element can be moved relative to a fixed slide to manipulate the fluid. In other embodiments, the slide is moved relative to a fixed opposable element to manipulate the fluid. In still other embodiments, both the slide and the opposable element are moved to manipulate the fluid. In addition, the two opposable elements are capable of processing a specimen. For example, two opposable elements can be used to attract and manipulate fluid to process a specimen held between the opposable elements. The specimen can then be transferred to a slide or suitable specimen carrier. Item 810 (fig. 59A and 59B) and opposable 2012 are non-limiting exemplary opposable elements and are discussed in detail in connection with fig. 64-67.

Fig. 64-67 illustrate one embodiment of opposable 810. Opposable 810 can include a body 1459, a port 1374, and a slot 1356. The body 1459 includes a first row of interstitial elements 1450, a second row of interstitial elements 1452, and a specimen processing region 1453. When the specimen processing region 1453 is oriented toward the slide and interfaced with a liquid, the liquid can be removed via the port 1374. The slots 1356 are capable of receiving features of an opposable actuator. The body 1459 can also include keying features 1362, 1364 (e.g., holes, protrusions, etc.) for aligning the opposable 810. The illustrated features 1362, 1364 are holes.

Fig. 64 shows a specimen processing region 1453 between two rows of gap elements 1450, 1452. Opposable 810 has edges 1454, 1456 that can be sized relative to the slide to provide a desired processing area 1453 (e.g., the entire surface 1460 of opposable 810, a majority of the upper surface 1460 of opposable 810, the area between interstitial elements 1450, 1452, etc.).

Fig. 65 shows an exemplary band of liquid 802 (illustrated in phantom) positioned between the gap elements 1450, 1452. The band of liquid 802 is able to move along the length of opposable 810 without contacting the gap elements 1450, 1452. The band of liquid 802 can be displaced without accumulating liquid around any of the gap elements 1450, 1452.

The gap elements 1450, 1452 can facilitate processing of the specimen with a desired amount of fluid (e.g., a minimum amount of fluid). The gap elements 1450, 1452 can also be spaced apart from one another to reduce, limit, or substantially prevent capillary action between adjacent elements. If the liquid 802 reaches one of the gap elements 1450, 1452, the liquid 802 can reside at the contact interface between that gap element and the slide without flowing to the adjacent gap element. The gap elements 1450, 1452 are spaced from the edges 1454, 1456 of the opposable 810 to maintain liquid proximate the treatment area 1453. Additionally, liquid 802 is held far enough away from edges 1454, 1456 to prevent wicking from below opposable 810 even if another object contacts edges 1454, 1456.

The rows of gap elements 1450, 1452 extend longitudinally along the length of opposable 810. The opposing gap elements of each row of gap elements 1450, 1452 are generally laterally aligned such that the slide can contact the laterally aligned gap elements 1450, 1452. As opposable 810 moves along the slide, the slide successively comes into contact with laterally aligned gap elements 1450, 1452.

Each row of clearance elements 1450, 1452 can be substantially similar to one another. Thus, the description of one of the rows of clearance elements 1450, 1452 applies equally to the other row, unless otherwise indicated. The row of gap elements 1450 can include about 5 gap elements to about 60 gap elements, wherein the average distance between adjacent gap elements is in the range of about 0.05 inches (1.27 mm) to about 0.6 inches (15.24 mm). In some embodiments, including the illustrated embodiment of fig. 64 and 65, the row of interstitial elements 1450 includes 19 interstitial elements that protrude outwardly from the entire surface 1460. In other embodiments, the row of gap elements 1450 comprises about 10 gap elements to about 40 gap elements. As seen from above (see fig. 65), the row of clearance elements 1450 has a substantially rectilinear configuration. In other embodiments, the rows of gap elements 1450 have a zig-zag (zig-zag) configuration, a serpentine (serpentine) configuration, or any other configuration or pattern.

The gap elements 1450 can be evenly or unevenly spaced from one another. The distance between adjacent gap elements 1450 can be greater than the height of the gap elements 1450 and/or less than the thickness T (fig. 67) of the body 1459 of the docking pod 810. Other spacing arrangements are possible, if needed or desired. In some embodiments, the thickness T is about 0.08 inches (2 mm). The width W between edges 1454, 1456 can be in the range of about 0.6 inches (15.24 mm) to about 1.5 inches (38 mm). In some embodiments, the width W is about 1.2 inches (30 mm), and the edges 1454, 1456 can be substantially parallel. Other widths are also possible.

Referring to fig. 65, the distance D between the rows 1450, 1452 can be selected based on the size of the specimen and the size of the slide. In some embodiments, distance D is in the range of about 0.25 inches (6.35 mm) to about 1 inch (25 mm). If the slide is a standard microscope slide, the distance D can be less than about 0.5 inches (12.7 mm).

Fig. 67 shows one of the gap elements 1450. The height H of the gap element 1450 can be selected based on the ability to manipulate the fluid. If the specimen is a tissue section having a thickness of less than about 0.0015 inches (0.038 mm), the clearance element 1450 can have a height H equal to or less than about 0.0015 inches (0.038 mm). If the gap element 1450 contacts the slide, the minimum height of the capillary gap (e.g., gap 930 of fig. 59A-63B) can be equal to 0.0015 inches (0.038 mm). In some embodiments, height H is in the range of about 0.001 inch (0.025 mm) to about 0.005 inch (0.127 mm). In particular embodiments, the height H is about 0.003 inches (0.076 mm) (e.g., 0.003 inches ± 0.0005 inches) to process thin tissue slices having a thickness of less than about 30 microns, about 20 microns, or about 10 microns.

The pattern, number, size and configuration of the interstitial elements 1450, 1452 can be selected based on the desired interaction between the specimen and the liquid. If the opposable 810 includes a field of interstitial elements, the interstitial elements can be uniformly or non-uniformly distributed across the opposable 810 to form different patterns that can include, but are not limited to, one or more rows, arrays, geometries, and the like.

The interstitial elements 1450 can be partially spherical dimples, partially elliptical dimples, or the like. The illustrated gap element 1450 is a substantially part-spherical dimple. If the specimen is large enough or moved toward one side of the slide, the gap elements 1450 in the form of dimples can slide over the specimen without damaging the specimen or bumping the specimen against the slide. In other embodiments, the clearance element 1450 can be in the form of a polyhedral protrusion, a conical protrusion, a frustoconical protrusion, or another combination of polygonal and arcuate shapes.

The body 1459 of fig. 66 is simple arcuate in shape having a radius of curvature R in the range of about 2 inches (5 cm) to about 30 inches (76 cm). In some embodiments, the radius of curvature R is about 15 inches (38 cm) or about 20 inches (74 cm). The nominal radius of the profile deviation can be equal to or less than about 0.1 inches. The actual radius of the profile can deviate by less than about 0.01 inches. Such an embodiment is well suited for producing a liquid band as follows: the liquid band has a generally rectangular shape when viewed from above and also spans the width of the slide, and has a low length variance along the slide for a particular volume. The radius of curvature R can be selected based on the number of specimens to be processed, the amount of fluid agitation, the properties of the processing liquid, the height of the gap elements 1450, 1452, and the like. In other embodiments, the opposable 810 can be shaped as a complex arc (e.g., an elliptical arc), a compound arc, and the like. In still other embodiments, opposable 810 can be substantially planar. The surface across the width W can be substantially flat.

Opposable 810 can be made in whole or in part of polymers, plastics, elastomers, composites, ceramics, glass, or metals, and any other material that is chemically compatible with the processing fluid and the specimen. Exemplary plastics include, but are not limited to, polyethylene (e.g., high density polyethylene, linear low density polyethylene, blends, etc.), polyvinylidene fluoride (PVDF), Polytetrafluoroethylene (PTFE), Perfluoroalkoxy (PFA), or combinations thereof. In some embodiments, opposable 810 can be made from a single material. In other embodiments, different portions of counter 810 may be made of different materials. If disposable 810, it can be made in whole or in part of relatively inexpensive materials. If the docking pod 810 can be rigid, it can be made in whole or in part of polycarbonate, urethane, polyester, metalized plate, or the like.

Referring again to fig. 66, the tip 952 includes an attracting feature in the form of a tapered region 1461. Tapered region 1461 is positioned to attract bands of liquid. As opposable 810 rolls over, the band of liquid can contact and cling to tapered region 1461. Curved surface 1463 provides a large surface area to which liquid can adhere. The illustrated tapered region 1461 has a radius of curvature equal to or less than about 0.08 inches to attract bands of liquid in cooperation with a standard microscope slide. Other radii of curvature can also be used if needed or desired. In some embodiments, the curvature of rounded edge 1461 is uniform across width W of opposable 810. In other embodiments, the curvature of the rounded edge varies across the width W of the opposable 810.

Opposable 810 can be disposable to prevent cross-contamination. As used herein, the term "disposable" is a broad term when applied to a system or component (or combination of components) such as opposable elements, treatment liquids, and the like, and generally means, but is not limited to, the system or component in question being used a limited number of times and then discarded. Some disposable components, such as the opposable element, are used only once and then discarded. In some embodiments, multiple components of the processing apparatus are disposable to further prevent or limit residual contamination. In other embodiments, these components are non-disposable and can be used any number of times. For example, non-disposable opposable elements can be subjected to different types of cleaning and/or sanitizing treatments without substantially changing the properties of the opposable elements.

The slides disclosed herein can be a 1 inch x3 inch microscope slide, a 25 mm x75 mm microscope slide, or another type of flat or substantially flat substrate. "substantially flat substrate" refers to, but is not limited to, any object having at least one substantially flat surface, but more generally refers to any object having two substantially flat surfaces on opposite sides of the object, and even more generally refers to any object having opposite substantially flat surfaces that are approximately equal in size but larger than any other surface on the object. In some embodiments, the substantially planar substrate can comprise any suitable material, including plastic, rubber, ceramic, glass, silicon, semiconductor materials, metals, combinations thereof, and the like. Non-limiting examples of substantially flat substrates include flat lids, SELDI and MALDI chips, silicon wafers, or other substantially flat objects having at least one substantially flat surface.

From the foregoing, it will be appreciated that specific embodiments of the invention have been described herein for purposes of illustration, but well-known structures and functions have not been shown or described in detail to avoid unnecessarily obscuring the description of at least some embodiments of the invention. The systems described herein are capable of performing a wide range of processes for preparing biological specimens for analysis. Where the context permits, singular or plural terms may also include the plural or singular term, respectively. Unless the term "or", "or" is expressly limited to one singular entity, excluding two or more other entities on the list, the use of "or" in this list may be understood to include: (a) any object on the list; (b) all objects on the list; or (c) any combination of the objects of the list. The singular forms "a", "an" and "the" include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to "a specimen" refers to one or more specimens, e.g., two or more specimens, three or more specimens, or four or more specimens.

The various embodiments described above can be combined to provide further embodiments. THE embodiments, features, systems, devices, materials, METHODS, AND techniques described herein may, in some embodiments, be compared to U.S. patent application No. 61/746,078 entitled "operation systems AND automatic speed processes SYSTEMS WITH operation systems" filed on 26/12/2012, U.S. patent application No. 61/746,085 entitled "automatic speed processes AND METHODS OF USING SAME" filed on 26/12/2012, U.S. patent application No. 61/746,085 filed on 26/12/2012, U.S. patent application No. 8298 entitled "speed processes SYSTEMS AND METHODS FOR modifying evaluation AND USING SAME", U.S. patent application No. 61/746,087 filed on 26/12/2012, U.S. patent application No. 2012 2 METHOD FOR program information pair information HEATING SLIDES "filed on 26/12/2012, U.S. patent application No. 61/746,089 filed on 4835/12/26, U.S. patent application No. 4935, SYSTEMS AND filed on 1/26, U.S. patent application No. 4935,3876 filed on 1/26/12/26, U.S. patent application No. 4935,, Any one or more of the embodiments, features, systems, devices, materials, methods, and techniques described in U.S. patent application No. 13/157,231, U.S. patent No. 7,468,161, and international application PCT/US2010/056752 are similar, all of which are incorporated herein by reference in their entirety. Furthermore, the embodiments, features, systems, devices, materials, methods, and techniques described herein may be adapted for use in or in conjunction with any one or more of the embodiments, features, systems, devices, materials, methods, and techniques disclosed in the aforementioned patents and applications in a particular embodiment. Aspects of the disclosed embodiments can be modified, if necessary, to employ concepts of the various patents, applications and publications to provide yet further embodiments. All applications listed above are incorporated herein by reference in their entirety.

These and other changes can be made to the embodiments in light of the above-detailed description. For example, the sealing element can have a one-piece or multi-piece construction and can include any number of retention features. In general, in the following claims, the terms used should not be construed to limit the claims to the specific embodiments disclosed in the specification and the claims, but should be construed to include all possible embodiments along with the full scope of equivalents to which such claims are entitled. Accordingly, the claims are not limited by the disclosure.

Claims (14)

1. An automated slide processing apparatus (100) for dispensing liquid onto one or more microscope slides (156), comprising:

a plurality of storage wells (180); and

a reagent pipette assembly (175) comprising a reagent pipette (204) movable between at least one loading position (213) for obtaining reagent from one of the reservoir wells (180) and at least one dispensing position for dispensing reagent onto one of the microscope slides (156);

characterized in that the automated slide treatment apparatus (100) further comprises:

a carousel (177) comprising the plurality of storage wells (180);

a wash pipette assembly (176) configured to wash the plurality of reservoir wells (180); and

a drive mechanism (184) coupled to the carousel (177) and configured to rotate the carousel (177) to position the reservoir wells (180) relative to the reagent pipette assemblies (175) and/or the wash pipette assemblies (176),

wherein the reagent pipette assembly (175) is movable between a filling position for obtaining reagent from a container (211) at a filling station (209) and a dispensing position for filling one or more of the reservoir wells (180) with reagent from the filling station (209).

2. The automated slide processing apparatus (100) of claim 1, wherein the filling station (209) comprises a plurality of containers (211) containing reagents; and wherein the automated slide processing apparatus (100) further comprises:

a plurality of slide treatment stations;

wherein the reagent pipette assembly (175) is movable through a lumen of the automated slide processing apparatus (100) to transport reagents obtained at the filling station (209) to the carousel (177) and dispense reagent mixture from the carousel (177) onto one of the microscope slides (156) at the slide processing station.

3. The automated slide processing apparatus (100) of claim 1, wherein the drive mechanism (184) is configured to sequentially rotate the reservoir well (180) under a wash pipette (213) of the wash pipette assembly (176).

4. The automated slide processing apparatus (100) of claim 1, wherein the reagent pipette assembly (175) is configured to fill the reagent pipette (204) with reagent from any of the reservoir wells (180).

5. The automated slide processing apparatus (100) of claim 1, further comprising a controller (144), the controller (144) communicatively coupled to the drive mechanism (184) and configured to command the drive mechanism (184) such that the drive mechanism (184) sequentially moves each of the reservoir wells (180) to a wash position for washing by the wash pipette assembly (176).

6. The automated slide processing apparatus (100) of claim 5, wherein the controller (144) stores and executes instructions for commanding the reagent pipette assembly (175) to fill the reservoir well (180) with reagent from a reagent container (211).

7. The automated slide processing apparatus (100) of claim 1, further comprising a controller (144) having mixing instructions executable to command the reagent pipette assembly (175) such that the reagent pipette assembly (175) delivers at least two reagents to one of the reservoir wells (180) to produce a reagent mixture.

8. The automated slide processing apparatus (100) of claim 7, wherein the controller (144) has mixed reagent dispensing instructions executable to command the reagent pipette assembly (175) to dispense the reagent mixture onto a specimen.

9. The automated slide processing apparatus (100) of claim 1, wherein the wash pipette assembly (176) is fluidly coupled to a vacuum source (237), and the wash pipette assembly (176) draws liquid from one of the reservoir wells (180) when the vacuum source (237) is evacuated.

10. The automated slide processing apparatus (100) of claim 1, wherein the carousel (177) includes spillways (187), the spillways (187) configured to allow reagent to flow from the storage wells (180) while preventing reagent from flowing between adjacent storage wells (180).

11. The automated slide processing apparatus (100) of claim 1, wherein the carousel (177) comprises a drain tube (183) and a plurality of spillways (187), the plurality of spillways (187) allowing overflow of reagents to flow from the storage well (180) toward the drain tube (183).

12. The automated slide processing apparatus (100) of claim 1, wherein the filling station (209) comprises a plurality of containers (211) containing reagents; and wherein the automated slide processing apparatus (100) further comprises: a slide treatment station, comprising:

a slide holder platen (601) having a receiving region (680) configured to receive a slide (156), wherein a first side of the slide (156) faces the receiving region (680) and a second side faces away from the receiving region (680); and

an opposable actuator (525) positioned to hold an opposable element (810, 2012) to define a capillary gap (930) between the opposable element (810, 2012) and a slide (156) located at the receiving region (680), the opposable actuator (525) configured to move the capillary gap (930) along the slide (156) in a first direction to move a band of liquid from a first position to a second position along a second side of the slide (156) and to reduce a width of the band of liquid in a direction substantially parallel to the first direction,

wherein the reagent pipette assembly (175) is movable through a lumen of the automated slide processing apparatus (100) to transport reagents obtained at the filling station (209) to the carousel (177) and dispense reagent mixtures from the carousel (177) onto microscope slides (156) at the slide processing station.

13. A method of dispensing a liquid onto a microscope slide (156), the method comprising:

sequentially delivering reagents to a plurality of storage wells (180) of a carousel (177) to produce a reagent mixture, wherein the carousel (177) is rotatable to sequentially position the storage wells (180) at a wash position;

at least partially filling a reagent pipette (204) with one of the reagent mixtures from one of the reservoir wells (180) when at least one of the reservoir wells (180) is at the purge location;

robotically moving the reagent pipette (204) toward the microscope slide (156) and dispensing the reagent onto the microscope slide (156) after at least partially filling the reagent pipette (204) with reagent; and

purging at least one of the storage wells (180) when the at least one storage well (180) is at the purge location,

wherein the reagent pipette assembly (175) is movable between a filling position for obtaining reagent from a container (211) at a filling station (209) and a dispensing position for filling one or more of the reservoir wells (180) with reagent from the filling station (209).

14. The method of claim 13, further comprising:

rotating the carousel (177) to position one of the storage wells (180) containing reagents at the wash position; and

washing the storage well (180) at the washing location to remove the reagent.