HK1118246A - Device with insert for analytical systems - Google Patents

Description

Device with insert for an analysis system

Technical Field

The present invention relates to a fluidic device for fluid analysis, said device having a first chamber and a second chamber and a channel leading from said first chamber to said second chamber, a method of using said device, an apparatus for analyzing fluids using said device and a system comprising said device and said apparatus.

The field of application of the fluidic device according to the invention is mainly fluid analysis, for example for analyzing nucleic acids in health care. The analysis performed using this device can be considerably improved as it avoids inaccuracies caused by contamination.

Background

In particular in analytical laboratories, there is great interest in performing analyses in a convenient, safe and reliable manner. A particular problem is the transfer from one reagent to the other. Devices for analysing samples and/or reagents have been proposed which minimise contamination of subsequent reagents in a continuous process.

In EP 318256 a device is shown comprising a chamber through which a fluid is forced. This device is not capable of performing more than one analysis.

In WO 93/22058 an apparatus is disclosed having a number of chambers each having a different temperature. The fluid flow in this device is complex.

It is an object of the present invention to provide a device with improved properties compared to devices according to the prior art, in particular a device allowing for simple fluid flow without transferring different reagents of a sample preparation step into a final measurement mixture, as in immunoassay and PCR-based amplification techniques.

Disclosure of Invention

A first subject of the invention is an analysis apparatus comprising a body comprising a fluidic unit comprising:

a) a first chamber having an outlet opening, an

b) A first passage leading away from said outlet opening,

the method is characterized in that: the first chamber further includes an insert including a second passageway, the insert being located in a first position in which the insert is not engaged with the outlet opening, the insert being movable from the first position to a second position in the first chamber in which the insert is engaged with the outlet opening such that the second passageway extends the first passageway into the first chamber.

A second subject of the invention is an analysis instrument comprising:

-a fitting for holding a device according to a general or preferred embodiment of the invention, and

-a head comprising an actuator, which reaches into the device with a degree of freedom to move the insert from said first position to said second position.

Another subject of the invention is a system for analyzing a fluid in a device, comprising:

-an apparatus according to a general or preferred embodiment of the invention, and

-an apparatus according to the invention.

Another subject of the invention is the use of a device according to a general or preferred embodiment of the invention for fluid analysis.

Yet another subject of the invention is a method for analyzing the composition of a fluid, the method comprising:

-providing a device according to a general or preferred embodiment of the invention or a system according to a general or preferred embodiment of the invention, and

-introducing a fluid into said first chamber,

-releasing said component of said fluid from other components of said fluid associated therewith in said first chamber,

-transferring the resulting fluid through said outlet opening and said first passage into said second chamber, said second chamber comprising a solid phase for immobilizing said component to be analyzed, thereby binding said component to said solid phase,

-moving said insert towards said outlet opening to said second position such that fluid may pass through said insert towards said outlet opening via said second passage and may pass through said recess, but may not enter said first passage when having entered said recess, and

-introducing a second fluid into said second chamber through said second channel.

Drawings

The first device according to the invention shown in fig. 1a is in a state in which the movable insert in the first chamber is in a first position, allowing fluid to freely exit from the chamber into the first channel through the channel in the insert and through the recess between the insert and the wall of the bottom part of the chamber. The device comprises 8 parallel analysis units, each equipped with an insert. The chamber and insert are shown in cross-section.

Figure 1b shows the same apparatus in a second position, allowing fluid to pass into the first channel only through the channel in the insert and not between the insert and the bottom of the chamber.

Fig. 2a shows an enlarged view of the insert in a first position in full view.

Fig. 2b shows an enlarged view of the insert in a first position in a sectional view.

Fig. 2c shows an enlarged view of the insert in the second position moved to the bottom of the chamber in a cross-sectional view.

Detailed Description

The apparatus of the invention is particularly useful in the analytical field for fluid action generally or desirably performed during fluid processing, such as physical processing and chemical processing of fluids. Because of the present invention, even complex fluidic processes can be performed. However, the present invention provides advantages even for simple steps. In an embodiment, more than one fluid may be processed in parallel.

The fluid that can be treated according to the present invention can be any fluid of interest that is subjected to a particular treatment. Preferably, the fluid is a liquid. More preferably, the liquid is an aqueous solution. In a preferred use of the device according to the invention, it is intended to analyze the composition of the liquid or the compounds obtained therefrom. In a diagnostic device, the liquid comprises a component to be determined in the analysis, such as a nucleic acid or an antigen. Such liquids may be selected from the group of: liquids from the environment, such as water from rivers or liquids extracted from soil, food fluids, such as fruit juices or extracts from plants or fruits, or fluids received from the human or animal body, such as blood, urine, cerebrospinal fluid or lymph, or liquids obtained therefrom, such as serum or plasma, or liquids comprising components to be separated from the aforementioned liquids. The liquid may further comprise additional components useful for the compositional analysis of the liquid or reagents for the chemical reaction to be performed within the apparatus. These reagents may include labeled binding partners, such as labeled oligonucleotide probes or dyes. Such agents are generally known to those skilled in the art.

The device according to the invention comprises at least one fluid unit. The fluid unit comprises at least one chamber and one first channel. In the following, a fluid unit will be understood as a configuration of chambers within a device, which chambers are interconnected such that a fluid introduced into one of said chambers may flow or may be caused to flow into another chamber of said configuration. In the simplest case, this is achieved, for example, by interconnecting the chambers and the channels so that fluid from the chambers can be forced into said channels. In case multiple analyses of different samples are to be performed within the same device, the device preferably comprises more than one fluidic unit. In this case, the fluidic behaviour of each fluidic unit is independent of the individual fluidic behaviour of the other fluidic units within the device.

Devices comprising chambers and channels are known. Devices having more than one chamber or/and more than one channel are also known. However, these prior art devices suffer from the problem that any fluid introduced through the same chamber after the first fluid is contaminated by the residue of the first fluid remaining in the chamber. The present invention addresses this problem, particularly for the case where the first chamber has a much larger volume than the subsequent second or third chamber, regardless of how many other chambers or channels are arranged in the fluid path behind the first chamber.

The manufacture of devices having at least one chamber and at least one passage from the chamber(s) is also known. Such a device can be easily prepared by injection molding methods using thermoplastic organic materials. In this case, any mold is configured such that the chambers and channels remain free of material through the molding process described. Other methods of machining a solid block of material to remove space for the chambers and channels, for example, may also be used.

The device according to the invention comprises at least one body. The body is a part of the entire device, which primarily provides rigidity or stiffness to the device. Thus, the body is preferably rigid. Preferably, the body is formed from a thermoplastic material, more preferably a material selected from thermoplastic organic polymers. Most preferably, the thermoplastic organic polymer is selected from the group consisting of polypropylene, polyethylene, polystyrene, polycarbonate and polymethylmethacrylate. It is further preferred that the material is light transmissive, at least at the portions required for the analysis. Depending on the amount and type of processing steps to be performed within the apparatus, the body may have a length of between 20 and 199mm, a width of between 8 and 30mm and a height of between 40 and 150 mm. Generally, the more fluid(s) to be analyzed, the larger the volume of the body.

A channel according to the invention is a cavity in the device whose longitudinal dimension is greater than its width and height. The channel is preferably bounded by walls defining the width and height of the channel. In a preferred embodiment, the channel walls are defined by the surface of a channel formed in the body of the device and the surface of the wall that is tightly sealed to the channel rim of the body. The channel formed in the device preferably has a diameter of less than 10mm²Preferably between 0.01 and 2mm²Cross section in between. The channel for transporting the fluid through the device will preferably have a smaller size than the chamber for holding the fluid or/and performing the process, which is preferably a chemical reaction.

The channels may have a variety of uses, for example:

-transporting a fluid between two locations (e.g. chambers) within the device,

-transporting a fluid into or out of the device,

measuring the fluid, or/and

-treating a fluid or treating a substance dissolved or suspended in a fluid.

A chamber according to the invention is another type of cavity within the device. The size of the cavity will vary depending on the intended use of the chamber. The chamber may have a variety of uses, for example:

storing, receiving or/and delivering a fluid, such as a sample or a reagent,

treating the fluid, e.g. for analysing substances within the fluid, or/and

measuring a physical or chemical property of the fluid (e.g. for performing optical absorption or fluorescence measurements).

The first chamber of the device of the invention is a chamber particularly suitable for an insert comprised in said chamber. As a first feature, the first chamber has an outlet opening. This opening is designed to allow fluid to leave the chamber. The fluid is then received by the channel exiting the chamber. The channel may be a channel as described above. The passage may lead to any other location within the device, but is preferably within the device, preferably a cavity within the device.

The channel is fitted in its shape to the outlet opening of the chamber so that it can receive fluid from said chamber through said opening.

According to the invention, the first chamber further comprises an insert comprising a second passage, the insert being in a first position wherein the insert is not engaged with the outlet opening, the insert being movable from the first position to a second position within the first chamber wherein the insert is engaged with the outlet opening such that the second passage extends the first passage into the first chamber.

Movement of the insert from the first position to the second position may be achieved by causing the insert to change its position within the device. This may be achieved by any force, for example mechanically, electrically or magnetically. The insert may slide along a path from a first position to a second position.

This arrangement of the insert in the second position allows the second fluid to be introduced into the first channel by personnel and/or instrumentation after the first fluid has selectively passed through the chamber without contaminating the second fluid with residues of the first fluid. Thus, a small remainder of the solution that has passed through said first channel cannot enter said first channel when urged to the bottom of the first chamber, for example by gravity or by vibration of the system, but is trapped in the now closed recess in the second position.

This is done by introducing a fluid into a channel in the insert leading to the first channel, i.e. the second channel. Any amount of the first fluid still present due to the previous processing step remains within the first chamber, in particular within the recess between the wall of the first chamber and the insert, since they cannot pass through the second channel extending into the chamber.

To achieve this, one or more, or even all, of the following measures may be taken.

In a first measure, the interior of the first chamber around the outlet opening area and the insert should be shaped to seal the connection between the first channel and the second channel. Preferably, the shape of said insert at the portion of the insert directed towards said outlet opening is similar to the shape of the first chamber. More preferably, the outlet opening widens conically from said first channel into said first chamber and said insert narrows conically, preferably at the same angle. This angle is preferably chosen between 5 and 85 degrees, more preferably between 20 and 60 degrees, with respect to the axis of outflow of the outlet opening, which may be in the same direction as the first passage out of the chamber.

The sealing area can be as small as 10mm²But preferably at 20 and 314mm²More preferably between 40 and 77.5mm²In the meantime.

In a second measure, the insert further comprises recesses between the ribs touching said first chamber wall. Thus, when the insert is in the first position, fluid may pass through the second passage and through the recess through the insert into the first passage via the outlet opening. On the other hand, when the insert is in the second position, fluid may pass through the insert via the second passage to the outlet opening and may pass through the recess, but may not enter the first passage when entering or/and passing through the recess.

In a third measure, the rib is located at a portion of the insert that touches the interior of the first chamber. The ribs provide for accurate positioning of the insert within the chamber. To improve this positioning, the part of the chamber in which the insert is movable and in contact with the rib has a diameter that is constant over the path of movement of the rib of the insert. This diameter is selected such that there is a constant pressure on the insert that is high enough to hold the insert in a defined position, but small enough to allow an operator to move the insert along a predetermined path of the insert. This can be achieved by using plastic material for the chamber walls and the inserts, in particular in the rib sections. The form of the mould giving the chamber its internal shape during injection moulding should be chosen to be slightly smaller than the form of the mould giving the rib its external form.

The form of the chamber portion in which the insert is guided by means of the ribs can have any form which allows the insert to move. Preferably, the diameter of the portion along the path of movement is simple, such as circular, rectangular or square. This is primarily because of the easier manufacturing, so any other form is also possible, but less preferred.

In an alternative embodiment, the walls of the chamber comprise the recesses and ribs described. In this case, the insert has a cylindrical form, the outer periphery of which is slightly larger than the inner diameter of the chamber.

In another embodiment, both the chamber and the insert have ribs and depressions. This is believed to improve the guiding feature of the chamber to the insert.

Preferably there are between 3 and 20, more preferably between 4 and 10 ribs around the periphery of the insert. There are depressions between the ribs. These recesses may have any form to allow the first fluid to be captured within said recesses. They may look similar or may be different.

In a first preferred embodiment, the outer shape of the insert is similar to the shape of the first chamber except for the top of the insert. Said top portion has a circular ring which increases conically from the outer boundary of the device towards the middle and tapers gradually from the inner second channel, so that an edge is formed on the top of the insert, which edge has a distance to said outer wall, so that the remaining fluid drop at the outer wall is caught between the edge and the outer wall, because the height of the drop is smaller than said distance.

In a further preferred embodiment, a further recess is formed between the outer wall of the insert and the inner wall of said first chamber, said inner wall forming at least 3, preferably up to 20 ribs to hold the body of the insert away from the inner wall of the first chamber and to reduce the force required to move said insert from its first position to its second position. The top of the insert is fitted with said conical ring, but this time the ring does not reach the inner wall of said first chamber, see fig. 1C.

The volume of fluid that cannot enter the outlet opening of the first chamber when the insert is in the second position is preferably between 10 and 500 μ l. This is sufficient to retain droplets of fluid from earlier processing that adhere to the walls of the first chamber and will contaminate the second fluid if they enter the fluid flow through the apparatus with subsequently added fluid.

The shape of the insert may be further influenced by the particular use of the insert. For example, it is preferred that the shape of the insert directed towards the inlet opening of the chamber comprises a tapered portion at the end of the second channel directed away from said outlet opening. This will allow better interaction of the insert with the actuator to move the insert in its path of movement towards the outlet opening. Furthermore, the tapered form may assist in directing the second fluid into the second channel.

The overall size of the insert may depend on the size of the first chamber. Preferably, the diameter of the insert will be no greater than the maximum diameter of the chamber, and if the insert is slightly larger in diameter, preferably 0.1 to 0.3mm, the insert is formed of a resilient material to ensure a desired fit of the insert within said first chamber. A preferred dimension for the diameter of the insert is between 0.1mm and 10cm, more preferably between 0.2mm and 20 mm. The length of the insert may be between 0.1mm and 10cm, more preferably between 0.1cm and 3 cm. The length of the rib along the path of movement may be between 0.1mm and 5cm, preferably between 0.2mm and 3 cm.

The inner wall of said first chamber is provided with at least one, preferably the same number of strips which hold the insert in said second position even when the force between the pipette tips moving the insert from said first position to said second position is greater than the sliding force with which said strips hold the insert in place. Thus, the pipette cannot pull the insert out of the second position, since the strip(s) will prevent the insert from moving back.

The length of the second channel is preferably between 0.1mm and 10cm, preferably between 1mm and 5 cm. Preferably, the channel is tubular, but may also include a tapered portion, preferably where designed for interaction with the actuator.

Preferably, the invention includes an actuator to move the insert within the device from the first position to the second position. This can be done by different means, either inside the device or outside the device. In a preferred embodiment, the actuator is a device comprised in an instrument for manipulating the device. Thus, the actuator may be independent in construction from the analysis apparatus. In this case, the device according to the invention preferably has an opening through which the actuator can access the interior of the first chamber to push the insert towards the outlet opening of the chamber. This opening may be an opening through which fluid is introduced into the chamber. Even more preferably, this opening is an upper opening of the chamber.

In a first use, the chamber will be used to receive a sample having a large volume, for example for performing a lysis reaction in the original sample, thereby adding a volume of reagent fluid. The volume of the chamber may be less than 1L, preferably between 1. mu.l and 100 ml. A preferred embodiment of such a chamber is a chamber for chemical sample preparation, e.g. lysis of cellular components of a fluid comprising cells, to release said cellular components, e.g. nucleic acids. Such a chamber may be referred to as a lysis chamber. The lysis chamber need not be a flat chamber, but will preferably have an at least partially tubular form, with an upper opening for introducing sample and reagents for lysis, and a lower opening as an outlet to the channel. The conditions under which chemical sample preparation is carried out are well known and readily applicable to the present invention.

In a preferred case, the fluidic unit according to the invention additionally comprises a second chamber. This chamber is fluidly connected to the first chamber by said first channel. The fluid unit may comprise even more channels and/or chambers, e.g. for further transporting the fluid to other chambers inside the device, or for further processing the fluid inside the device.

In a preferred embodiment, it is useful for determining nucleic acid analytes within a fluid, that each fluidic unit comprises a first chamber, i.e. a lysis chamber, and a first channel leading from an outlet portion of said first chamber, preferably through an inlet portion, to a preferably flat second chamber comprising a down-let (fleece) capable of reversibly binding nucleic acids. A third channel leading from the outlet portion of said second chamber, preferably through the inlet portion of the third chamber, to said third chamber, and a fourth channel leading from the outlet portion of said third chamber to the outlet of said device. Any of these chambers may be a chamber according to the present invention. Preferably, the chamber defined according to the invention is a lysis chamber and is a first chamber according to the invention.

More preferably, the fluid unit further comprises a third channel leading from said second chamber to a third chamber for illumination and detection.

The last channel in the fluid path, i.e. the fourth channel in the above embodiments, exits the device through an outlet. The outlet in the device according to the invention, more preferably the outlet of the fluid unit, is an opening of the device, which is designed to allow the fluid to leave the device in a controlled manner, while avoiding unintended escape of the fluid during treatment. Preferably, therefore, the opening is sealed, for example by a plug pierceable by a hollow needle.

The device according to the invention may comprise as many fluid units as are meaningful. An excessive number may be disadvantageous in view of more difficult handling of the device. For example, this may require too many actuators within the instrument to fluidly accommodate. It has proven advantageous to use from 2 to 16 fluid cells, more preferably from 4 to 8 cells, in one device.

For convenient handling of the apparatus, the fluid cells are preferably arranged in a parallel pattern. This means that the positions of the chambers and channels of the different fluid cells are geometrically parallel to each other. Any inlets and outlets are then located at the same side of the device, preferably each type of port, e.g. inlet, is located along an edge of the device, and the other types, e.g. outlets, are located along another edge. If there are two different types of inlets, they may be arranged on the same side or edge of the device.

The form and size of the entire device according to the invention is mainly determined by the function the device is to be used for. Furthermore, the type and amount of fluid in the device and the type and number of steps to be performed further determine the geometric and functional characteristics of the device.

Fluidic devices according to the understanding used herein have one or more channels with dimensions greater than 0.1 μm²More preferably at 10 μm²To 10mm²Cross section in between. The device may further or alternatively comprise one or more chambers having a larger cross-section than the channel. The chamber of the fluidic device may have a volume of between 10 μ l and 3ml, more preferably between 1 μ l and 5 ml.

The device according to the invention may comprise further elements such as recesses and protrusions for interacting with instruments for receiving and/or handling said device. Preferably the apparatus includes a channel to engage with the clamp to grip the apparatus and transport the apparatus to a position within the instrument and secure it in a predetermined relative position.

A first preferred embodiment of the device according to the invention is shown in fig. 1 a. The device 1 according to the invention is shown in a state in which the movable insert 2 in the first chamber 3 is in the first position, allowing fluid to freely leave from the chamber, through the second channel 5 in the insert 2 and through the recess between the ribs of the insert and the wall of the bottom part/outlet part 4 of the chamber 3 into the first channel 6. The ribs and depressions of this case are shown in more detail in fig. 2 b. The device comprises 8 parallel analysis units, each equipped with an insert. The chamber and insert are shown in cross-section.

Fig. 1b shows the same device 1 in a second position, allowing fluid to exit into the first channel 6 only through the channel 5 in the insert 2, and not between the insert 2 and the bottom 4 of the chamber 3.

Fig. 2a shows an enlargement of the lower part of the first chamber with the insert 2 in the first position, the insert being shown in a 3D view, while fig. 2b shows the same in a cross-sectional view and fig. 2c shows the insert in the second position. In each figure are shown a first channel 6, a bottom part 4, a recess 8, a rib 7 and a second channel (only visible in the cross-sectional view), in this case 6 recesses around the periphery of the insert 2 and some hidden.

More preferably, the device according to the invention is a composite of a body and at least one sealing wall. In this case, any cavity within the body is closed by a sealing wall attached to the body, except for the cross-section of the passage through which fluid may enter or/and exit the chamber and the inlet and outlet of the device.

Preferably, the body has a generally flat region with an area of 1600 to 19200mm²More preferably between 7200 and 12000mm²In the meantime. This region is hereinafter referred to as the sealing region. The term "flat" means that the body is geometrically homogeneous towards the outside of the device to allow the sealing unit to closely and thermally contact the body so that sufficient heat can be applied to the material of the body to melt the portion of the body in contact with the sealing unit. In other portions, the body may include a region raised from a flat surface, for example, near a chamber formed within the body.

The sealing wall is preferably a generally flat piece of material. It may be made of one material or may be a composite. Preferably, it has the form of a foil which is less rigid than the body. The present inventors have found that it is very advantageous if the sealing wall is a composite of the same thermoplastic material as the body-this part being referred to as the thermoplastic part-and a carrier part made of a material having a melting temperature higher than the melting temperature of the thermoplastic part. Preferably, the carrier portion is selected to provide tear strength to the closure wall. The tear strength is important for the reliability of the sealing process. Preferred tear strength is preferably in the range of 5 to 50N/mm²More preferably between 6 and 40N/mm²In the meantime. Preferred materials for the carrier part are selected from the group of metal foils; more preferably the material comprises aluminium. The thickness of the foil is preferably between 40 and 400 μm.

Preferably, the sealing wall is a heat transfer wall. The heat transfer wall preferably comprises a heat transfer material, i.e. a material having a good thermal conductivity. The preferred heat transfer material is selected from the group of aluminum and copper, more preferably aluminum. Preferably, the heat transfer wall comprises 2 layers, preferably one of said layers is a metal layer and the second layer is a thermoplastic layer, and said layers are welded together.

To ensure a precise seal, in particular a liquid-tight seal, of the sealing wall to the body in the region around the cavity, the sealing wall is preferably substantially flat. By substantially flat is meant that the sealing wall is flat over more than 80% of its surface, preferably over more than 90% of its surface and most preferably over 100% of its surface. The portion of the body intended to be sealed to the sealing wall should be substantially flat to similarly extend within the region surrounding the cavity, but not include the groove intended to form the channel or chamber within the device after sealing.

The thickness of the sealing wall is preferably between 20 and 1000 μm, more preferably between 50 and 250 μm. Preferably, each body of the device has a sealing closure, covering all the channels to be sealed in the body.

In said fluid cell, at least one cavity is formed between these components. The cavity includes at least one chamber and at least one channel. The fluidic unit according to the invention thus requires at least one chamber, referred to as first chamber, and at least one channel, referred to as first channel. This fluid unit is located at the location of the device in question, for example at the beginning of the fluid path, which is accessible by the actuator from outside the device, so that the actuator can access the interior of the chamber.

The two parts, the body and the sealing wall, can be joined by known methods. In a preferred embodiment, where the sealing wall is a thin wall comprising a thermoplastic polymer and the rigid body is made of a polymer, such as polystyrene, the two parts can be combined and then sealed by welding, such as by laser welding, ultrasonic welding, heat sealing or gluing. The two parts may also be merely clamped or glued together.

The bonding method, the material of the body and the material of the sealing wall must be chosen to cooperate with each other. For example, if the bonding method is laser welding, the bulk material of the body and the sealing wall is the same material (e.g., polypropylene), but one of the two materials is dyed to have absorption for the laser energy. If the bonding method is ultrasonic welding, the two materials are typically the same. If the bonding method is heat sealing, the sealing wall is a heat sealable wall adapted to be heat sealed to the body.

In the above method for manufacturing, further assembly steps may be added, in particular if the device comprises further elements.

Another subject of the invention is an apparatus comprising:

a fitting for a holding device according to the invention or a preferred embodiment thereof, and

-a head comprising an actuator, which reaches into the device and has the freedom to move the insert from said first position to said second position.

In order to hold and apply the instrument arrangement reliably to the device, the instrument comprises a fitting for holding the device. This fitting also allows to keep the device in a position where fluid can be introduced into the fluid unit of said device at the desired time. The fitting may be adapted as well as possible to the external form of the device in question to hold the device. The mating member may comprise snap-in means in the form of parts of the mating device. Such a form fit may be provided by a protrusion in the fitting which is insertable into a recess in the device, or vice versa.

Furthermore, the instrument according to the invention comprises a head comprising an actuator reaching into the device and having a degree of freedom to move the insert from said first position to said second position. In the sense of the present invention, the actuator is a device having rigidity to push the insert from the first position to the second position. The insert does not need to be moved back by pulling force. In a preferred embodiment, the inner wall of said first chamber has a strip which holds the insert in said second position. A preferred actuator according to the present invention is a pipette tip mounted to a mount on the pipette manager. This has the advantage that no additional equipment is required for moving the insert. In addition, no time-consuming equipment changes are required. The insert can simply be pushed towards the outlet opening using the pipette tip used in this pipetting step, which is preferably the first dispensing step of the washing procedure. Such pipette tips and pipette managers are generally known to those skilled in the art. The only modification compared to the instruments currently used in laboratories is to adjust the end position of the pipette tip exactly to the second position. The computer program controlling the process steps needs to be adjusted to an additional forward movement to push the insert towards the second position. No additional building blocks are required for existing instruments.

An instrument according to the invention may comprise a device to fulfil a dispensing or delivery function, and removal or receipt of fluid to and from the device should be considered as both active and passive manipulation. For example, receiving fluid from the first fluid handling unit may be accomplished by applying fluid under pressure to the apparatus to press the fluid into the apparatus or by applying negative pressure to the cavity to draw the fluid into the apparatus, and removing or transporting fluid from or to the outside of the apparatus may be accomplished by applying pressure to the cavity, for example by pumping fluid such as liquid or gas through the first inlet, or by applying negative pressure to the cavity to draw the fluid through the inlet. Suitable devices include syringe pumps. The liquid handling units are located within the instrument such that they can function in any input and output position when the device is put into a defined position on the instrument. The position of the head relative to the inlet or outlet of the apparatus may be controlled by the control unit.

Preferably, the instrument is an analytical instrument. Instruments for analyzing a fluid or any component of a fluid are generally known. They include units generally known for analysis. Preferred units are optics for determining a property, e.g. an optical property or a change in a property, of a fluid comprised in the device, a machine for moving a fluid from a first location to one or more other locations, and a liquid handling unit for dispensing or/and extracting a fluid from a tube, vessel or reagent container into the device. As indicated above, the instrument comprises a head for dispensing a fluid into a fluid unit of the device according to the invention, or/and removing a liquid from the device.

The apparatus further preferably comprises a heater, preferably a heating or/and cooling element. This element is positioned such that, preferably when the fluid is comprised in a chamber within the device, it contacts or can contact the device at the outside of the sealing wall, such that heat can be transferred from the chamber to the heater or/and cooler and from the heater or/and cooler to the chamber, preferably through said heat transfer wall. An example of an instrument that includes heating or/and cooling elements is a thermal cycler. Thermocyclers are generally known to apply different temperature profiles to a fluid in a repetitive manner. A typical thermocycler is described in EP 0236069. Preferred heating or/and cooling elements are selected from the group comprising peltier elements, resistive heating elements and passive cooling elements such as metal blocks equipped with fans.

In the present invention, there is preferably at least one thermal cycler unit for each fluid unit, each thermal cycler unit being located in a position within the instrument that is moved relative to the apparatus to contact said sealing wall close to the chamber containing the fluid to be heated. Preferably, this chamber is a third chamber as indicated above. More preferably, each thermal cycler is independently adjustable, i.e., each thermal cycler can be applied with a different thermal profile. The thermal profile is defined by the temperature to be reached in the chamber and the length of time to maintain this temperature. The different profiles may be implemented by computer control. The provision of a break to the device facilitates the ability to use different thermal profiles at adjacent fluid cells.

To monitor a property or a specific change of the liquid during a process performed in the apparatus, the instrument further comprises a property monitor unit, e.g. a detection module, optically connected to the transparent wall of the chamber in said fluid unit. Suitable detection modules are generally known and depend on the type of property or the type of change in property that is made during the presence of fluid within the device. For example, if the characteristic is a change in an optical signal, e.g. a fluorescence signal, the detection module will comprise a light source positioned within the device such that a fluid cell of the device, preferably a detection chamber within the device for this purpose, e.g. a fourth chamber, can be illuminated, and an illumination receiving unit, preferably a light sensitive element for receiving illumination from the fluid comprised in the device and transferring an electrical signal to the evaluation unit. The detection module is located within the instrument at a position such that it can detect light emitted from the fluid contained within the chamber. It is preferred if there is also an illumination module positioned to project light into the room; this light preferably has characteristics that excite components or are absorbed or altered within the fluid.

If the process being performed within the device requires that components of the device, such as electrodes or heated walls within the device, be connected to the circuitry of the instrument, such connectors are preferably provided at locations on the instrument that cause the connectors on the instrument to connect to their counterparts on the device when the device is inserted into the instrument.

Another subject of the invention is a system for analyzing a fluid in a device, the system comprising:

-a device according to the invention in its general or preferred embodiments; and

the apparatus according to the invention in its general or preferred embodiment.

Preferably, the system according to the invention comprises a further fluid container (e.g. for waste collection) or/and one or more reagent containers.

A further subject of the present invention is the use of a device according to the present invention in its general and preferred embodiments in a method for analyzing a sample.

Thus, another subject of the invention is a method for analyzing the composition of more than one fluid, the method comprising:

-providing a device according to the invention or a preferred embodiment thereof or a system according to the invention or a preferred embodiment thereof, and

-introducing a fluid into said first chamber,

-releasing said component of said fluid from its associated other component of said fluid in said first chamber;

-transferring the resulting fluid through said outlet opening and said first passage into said second chamber, said second chamber comprising a solid phase for immobilizing said component to be analyzed, thereby binding said component to said solid phase,

-moving said insert towards said outlet opening to said second position such that fluid may pass through said insert towards said outlet opening via said second passage and may pass through said recess, but may not enter said first passage when having passed through said recess,

-introducing a second fluid into said second chamber through said second channel.

The second fluid may be introduced through the second channel and the first channel, since in the second position the second channel is directly connected to the first channel.

The fluid, preferably the sample or/and the reagent to be analyzed, may be introduced into the apparatus according to known methods, e.g. by pipetting the fluid into an opening in the fluidic unit. Preferably, the fluid is introduced into the fluid unit through a head for an instrument as outlined above, for example using a pipette tip carried by the head through said inlet into the first chamber. Within these chambers, the sample is processed to release the components of the sample to be analyzed from any cell chambers within the sample to which the components may be associated. For analysis of nucleic acids, this may include a combination of chemical treatment to digest the cell wall with chaotropic salts and proteases and physical treatment to destroy the cells by heating, for example by heating the lysis mixture to between 37 ℃ and 38 ℃. The exact conditions may depend on the particular type of sample and lysis solution and/or enzyme used for lysis. Some samples may require more harsh conditions than others. To achieve lysis, the sample must be contacted with a reagent for processing, e.g., for lysis. This is preferably done by pipetting an aliquot (aliquot) of each of the sample and reagent into the chamber.

If sample preparation is intended solely within the device, the process is completed by removing the pre-treated sample from the chamber, e.g. by removing the mixture through the first channel. However, other steps may be added to the described apparatus, which may or may not include further embodiments of the invention.

If the method according to the invention shall perform an analysis comprised within the sample, the method according to the invention shall comprise, after the treatment in the first chamber, e.g. after lysis of the sample, transporting the result of the step, e.g. the pre-treated sample, into the second chamber for further processing. This is preferably done by subjecting the fluid to a positive or negative pressure leaving the first chamber through the outlet portion into the first passage. In a preferred embodiment, a fluid for purification purposes that is a component of the sample is transferred into the second chamber. Any components to be immobilized are bound to the porous material included therein.

A particularly preferred embodiment of the invention comprises introducing a second fluid into said second chamber through said second passage after the insert has been moved to the second position, preferably by means of the actuator outlined above. Preferably, said second fluid is selected from the group comprising a washing fluid and/or an elution buffer and/or a master mix (master mix). A wash buffer is a fluid designed to remove any free components of the fluid from the component(s) immobilized to the porous material. Such buffers are well known in the art and preferably comprise a lower salt concentration than the fluid used for immobilization. The elution buffer preferably comprises reagents for detecting components of or derived from said fluid. The mixture of elution buffer and master mix further includes reagents for amplifying and detecting nucleic acids, such as primers, probes, enzymes, and reagents.

To perform the assay, the method preferably comprises first rinsing the components immobilised on the porous material and then eluting them from the material.

The eluate is then preferably passed to a third chamber for detection. This may be done by supplying fluid to the apparatus, preferably through said second channel. This will force fluid through the third channel to the third chamber.

The chamber preferably further comprises an outlet portion for the fourth channel at its end opposite the inlet portion of the third channel, said fifth channel leading to another fluid port, i.e. the outlet.

In a preferred embodiment, there is at least one chamber within each fluid unit, more preferably a third chamber as outlined above, designed to allow a step of physical or chemical treatment of said fluid. Preferably, the physical treatment is a treatment selected from the group of heating and cooling (thermal treatment), mixing and irradiation, and any combination thereof. Any heat treatment may be performed through any wall of the chamber of the apparatus. Preferably, the heating is done through a sealing wall.

In a first preferred embodiment, the physical treatment is thermal cycling as used in polymerase chain reaction (PCR, EP 0201184).

In another preferred embodiment, the first or third chamber in each fluidic unit is preferably a detection chamber, and most preferably an amplification/detection chamber. Within this chamber, a measure is preferably determined which represents the component to be analysed or which represents a property of the component derived from the component to be analysed as being the presence or absence of the amount of the component of the original liquid.

The detection may be a two-step process comprising illumination and monitoring. After irradiating the fluid in said chamber, the contents of the chamber, i.e. the properties of the fluid, are monitored. The monitoring of the fluid property may be through a wall of the body. The requirements of the monitoring process determine the characteristics of the walls that bound the chamber. For example, determining light emanating from the fluid using a detector unit located outside the device within the instrument requires that the walls be transparent to the light emanating from the chamber. In this case, the material of the wall will be a material transparent to this light. If said monitoring additionally requires that light is projected through said wall onto the fluid comprised in said chamber, the material of the wall should be transparent for the projected light.

Detection may be accomplished by illuminating the liquid in the chamber with light having a wavelength at which one of the components or agents in the fluid has a measurable absorption. Determination of the light leaving the cavity, for example by fluorescence, may be used to determine the absorbance of the liquid or any change in the absorbance of the liquid over time or compared to a standard liquid.

The chemical treatment is the performance of a chemical reaction. Preferably, in the third chamber, the progress of the chemical reaction is detected. A preferred chemical reaction is one that modifies the chemical composition of any component of the fluid or any derivative thereof. More preferably, the chemical reaction is selected from the group consisting of primer extension, hybridization, denaturation and cleavage. Most preferably, the chemical reaction is a PCR as referred to above or a modification thereof, e.g. a homogeneous PCR, sometimes also referred to as a real-time PCR, as described in EP 0543942. In real-time PCR, the signal is not necessarily determined at the end of the amplification reaction, but at least once between the first and last thermal cycles.

To perform combined amplification/detection including PCR, the contents of the chamber are heated and cooled in a cyclic manner. To achieve efficient thermal cycling, the containment wall covering the third chamber includes a metal portion, i.e., a heat transfer foil, that facilitates heat transfer from the thermal cycler into the chamber. Homogeneous PCR allows detection through a transparent window in the body almost from the beginning of thermal cycling.

In a very preferred embodiment of this analysis method, the component of the liquid to be analyzed is a nucleic acid suspected to be comprised in the fluid, for example a part of the hepatitis c virus genome. The reagents for the assay, preferably elution buffers, will then include reagents, such as primers, for amplification of the particular segment of the nucleic acid in question, and also probes for binding to the amplified segment. A very preferred example of such a reaction is disclosed in EP 0543942. To apply thermal cycling to the fluid contained within the chamber, the instrument used includes a combined heating/cooling block to bring the contents of the chamber to a temperature within the curve (profile) required for nucleic acid amplification. The change in absorbance or fluorescence within the fluid is then used as a measure of the nucleic acid to be determined within the fluid.

The reagents for the processes within different fluidic units of one device may be the same or may be different. For example, if HBV is to be detected in a first fluidic unit and HIV is to be detected in a second fluidic unit, the same procedures and reagents used for sample lysis and purification can be used for both aliquots of the sample in different units, but different reagents should be used for amplification and detection (elution buffer and master mix), thus reflecting the different order to be amplified. Suitable reagents for sequence-specific amplification and detection are known to those skilled in the art and may be similarly applied.

The preferred embodiments have been detailed above in the description of the apparatus according to the invention.

The advantage of the device according to the invention is that the device avoids contamination of the fluid subsequently used in the device in a simple manner. Furthermore, in a preferred embodiment, several analyses may be performed in parallel, even if the analyses differ, for example in determining different analytes or in performing different chemical reactions.

Reference numerals

1 apparatus according to the invention

2 insert

3 first chamber

4 outlet part

5 second channel

6 first channel

7 Ribs

8 depressions

9 second chamber

While the foregoing invention has been described in some detail for purposes of clarity and understanding, it will be clear to one skilled in the art from a reading of this disclosure that various changes in form and detail can be made without departing from the true scope of the invention. For example, all of the techniques and instruments described above may be used in various combinations. All publications, patents, patent applications, and/or other documents cited in this application are incorporated by reference in their entirety for all purposes to the same extent as if each individual publication, patent application, and/or other document were individually indicated to be incorporated by reference for all purposes.

Examples of the present invention

Example 1

Manufacture of the device according to the invention

a) The device as shown in fig. 1a is prepared as follows:

two-part moulds, which reflect the external form of the apparatus according to FIG. 1a, are filled with polypropylene (Handbuch Spritzgie. beta. en, 2004, Hanser Verlag, p. 77) (Werkstoff-F ü hr Kunststoffe, 2001, 8. autoflage Hanser Verlag, p. 83-89).

The large chamber has a tubular section (diameter: 14mm, height: 40mm) in its lower part. The angle at the bottom is 65 degrees. After curing, the foils of polypropylene (30 μm) and aluminium (110 μm) were welded to the polypropylene body with a heat weld. The outlet opening is closed by a silicon plug.

b) The insert shown in fig. 1a-c is produced from a slightly elastic polymer by injection moulding (diameter of the ribs: 14mm, rib height: 12mm, channel width: 0.8mm, lower angle same as the apparatus). The insert is inserted into the upper part of the tubular part of the chamber of the device manufactured by a).

c) The fiberglass fluff is inserted into the second chamber. The device is then sealed with a sealing foil (heat transfer foil).

Example 2

The procedure comprising sample preparation and PCR and detection carried out in one of the devices of example 1a was carried out

In a first step, the device as manufactured in example 1 is loaded into a processing station of an instrument. This is done by using a clamp (see fig. 1a-c, reference numeral 8) which engages into a recess on the upper part of the device. Then, a quantitative standard solution was added to each of the first chambers using a head supporting 8 pipette tips (see 3). The tip is discarded. The lysis solution comprising proteinase K was then added to the chamber again using parallel pipetting. Then a new pipette tip was used to add 7 samples and one negative control aliquot to each of the first chambers. The tip serves to thoroughly mix the solution by aspirating and spitting the mixture in each chamber. The mixture was then incubated at 72 ℃ for 10 minutes to lyse while the tip remained in the first chamber.

Pressure is then applied to the system to transport the mixture through the outlet portion of the first chamber (see 4) to the second chamber filled with glass wool (see 5). Any nucleic acid binds to the glass surface and liquid is removed through the outlet. A hollow steel needle is docked to the outlet to withdraw the liquid. The pipette tip is then moved downward until the lower tapered portion of the insert touches the tapered portion of the bottom of the chamber of the device. Thus, the insert is pushed towards the bottom, thus leaving only the passage in the insert for the liquid to pass through the outlet opening of the chamber. Several drops of fluid from the wall of the lysis chamber move from the wall to the bottom and are retained within the recess of the insert.

After removal of the pipette tips, a rinsing liquid (400 μ Ι) was pipetted into the conical part of the insert using a new set of pipette tips and sucked through the second chamber, thus removing impurities from binding with the nucleic acids. This was repeated three times.

An aliquot (70 μ l) of elution buffer is added to the tapered portion of the insert in the first chamber and pipetted through the fluidic unit so that the eluted liquid remains in the third chamber.

The liquid in the third chamber is subjected to a thermal cycle as follows

First cycle

50 ℃, 120 seconds, UNG step

5 cycles

Denaturation at +4 ℃/sec, 95 ℃ for 15 sec

Annealing at 59 ℃ 4 ℃/s for 50 s and fluorescence measurement after 35 s

45 cycles

Denaturation at +4 ℃/sec, 91 ℃ for 15 sec

4 ℃/sec 50 sec annealing at 52 ℃ and fluorescence measurement after 35 sec

Light having a wavelength of the excitation wavelength of the probe is projected into each third chamber and fluorescence is measured in the third chamber during illumination during the annealing phase of each cycle. The amount of nucleic acid in each sample was determined by standard calculations using quantitative standards. The eighth sample serves as a negative check.

Claims (16)

1. An analytical device comprising a body including a fluidic unit comprising:

a) a first chamber having an outlet opening, an

b) A first passage leading away from said outlet opening,

the method is characterized in that: the first chamber further comprises an insert comprising recesses between ribs touching the wall of the first chamber and a second channel, the insert being in a first position in which the insert is not engaged with the outlet opening, the insert being movable from the first position to a second position in the first chamber in which the insert is engaged with the outlet opening such that the second channel extends the first channel into the first chamber and resembles the shape of the chamber in the part directed towards the outlet opening.

2. The apparatus of claim 1 wherein said outlet opening widens conically from said first passage into said first chamber.

3. The apparatus of claim 1 or 2, wherein when said insert is in said first position, fluid can pass through said second passageway and through said recess of said insert and through said outlet opening into said first passageway.

4. Apparatus according to any preceding claim, wherein when said insert is in said second position, fluid can pass through said insert to said outlet opening via said second passage and can pass through said recess, but cannot enter said first passage when having passed through said recess.

5. The apparatus of any of the preceding claims, wherein the insert further comprises a tapered portion at an end of the second channel directed away from the outlet opening.

6. Apparatus according to any preceding claim, wherein the volume of fluid unable to enter the outlet opening of the first chamber when the insert is in the second position is between 5 and 1000 μ l.

7. An analytical instrument comprising

-a fitting for holding a device according to any of claims 1 to 6,

-a head comprising an actuator, which reaches into the device and has a degree of freedom to move the insert from said first position to said second position.

8. The instrument of claim 7, further comprising two or more pipette tips mounted on the mount, the pipette tips having an outlet opening with an external conical shape.

9. A system for analyzing a fluid within a device, comprising

-a device according to any of claims 1 to 6, and

-an apparatus according to any of claims 7 to 8.

10. The system of claim 9, further comprising a fluid dispensing unit.

11. Use of the device according to any of claims 1 to 6 for fluid analysis.

12. A method of analyzing a fluid composition comprising

-providing a device according to any of claims 1 to 6 or a system according to any of claims 8 to 9, and

-introducing a fluid into said first chamber,

-releasing said component of said fluid from other components of said fluid associated with the component in said first chamber,

-transferring the resulting fluid through said outlet opening and said first passage into said second chamber, said second chamber comprising a solid phase for immobilizing said component to be analyzed, thereby binding said component to said solid phase,

-moving said insert towards said outlet opening to said second position such that fluid can pass through said insert towards said outlet opening via said second channel and can be captured in said recess but cannot enter said first channel when captured in said recess,

-introducing a second fluid into said second chamber through said second channel.

13. The method of claim 12 wherein said second fluid is introduced through a pipette tip docked in a fluid-tight manner in said second channel of said insert.

14. The method of any of claims 12 or 13, wherein the second fluid is a wash buffer.

15. The method of any of claims 12 to 14, further comprising:

-introducing a third fluid into said second chamber through said second passage.

16. The method of any of claims 12 to 15, further comprising removing said third fluid from said second chamber into a third chamber with said component,

-heat treating said third fluid comprising said ingredients in said third chamber.