JP2007003350A - Perforating implement for dispensing device - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To perforate at low cost an evaporation prevention film for covering an opening part of each well. <P>SOLUTION: A pipette chip for sucking solution from each well is mounted exchangeably on a nozzle 40 of a dispensing head 87. A perforating pin 20 is mounted by utilizing the nozzle 40. The perforating pin 20 is constituted of an adapter 65 and a peak part 66. The adapter 65 is formed by notching the tip of the pipette chip, and a cap 67 pressed into the tip 68 of the nozzle 40 is provided on the rear end. The peak part 66 is constituted of a screwing part 80 inserted into an opening on the tip 69 of the adapter 65, a flange 70 abutting on the tip 69 of the adapter 65, and a pin 75 for breaking the film, and fixed to the adapter 65 by screwing a bolt 94 inserted from the inside on the cap 67 side into a screw hole 102 provided on the screwing part 80. <P>COPYRIGHT: (C)2007,JPO&INPIT

Description

  The present invention relates to a hole punch used in a dispensing device having a dispensing head to which a pipette tip for sucking a liquid is attached, and more specifically, before aspirating a liquid contained in each well of a well plate, The present invention relates to a punch for breaking a film covering the opening of each well.

  For example, a measurement device using total reflection attenuation is known as a measurement device for measuring the reaction of a sample when investigating the interaction of biochemical substances such as proteins and DNA, or when performing drug screening.

  A measuring device using total reflection attenuation causes a sample reaction on the sensor surface, which is one surface of a thin film formed on a transparent dielectric, and causes a total reflection condition on the light incident surface on the back surface of the sensor surface. The reaction is measured by making light incident so as to satisfy the above condition and detecting the attenuation of the reflected light. One of the measuring devices using such total reflection attenuation is a measuring device using a surface plasmon resonance phenomenon (hereinafter referred to as an SPR measuring device). The surface plasmon is a density wave of free electrons generated by collective vibration of free electrons in a metal and traveling along the surface of the metal.

  The SPR measurement device uses a metal film as a thin film formed on a transparent dielectric, and uses one surface of the metal film as a sensor surface to generate SPR on the sensor surface, and the reaction state of substances generated there Measure by detecting SPR.

  When light is irradiated from the back side of the sensor surface of the metal film so as to satisfy the total reflection condition (at an incident angle greater than the critical angle), total reflection occurs at the light incident surface, but only a small amount of incident light is It passes through the metal film without reflection and oozes out to the sensor surface. This light wave that oozes out is called an evanescent wave. When the frequencies of the evanescent wave and the surface plasmon coincide and resonate (when SPR occurs), the intensity of the reflected light is greatly attenuated. The SPR measurement device detects the SPR generated on the sensor surface on the back side by capturing the attenuation of the reflected light reflected by the light incident surface.

  The incident angle (resonance angle) of light for generating SPR depends on the refractive index of the medium through which the evanescent wave and the surface plasmon propagate. In other words, if the refractive index of the medium changes, the resonance angle that generates SPR changes. The substance in contact with the sensor surface becomes a medium for propagating evanescent waves and surface plasmons. For example, when a chemical reaction such as bonding or dissociation between two types of molecules occurs on the sensor surface, it is determined by the refractive index of the medium. It appears as a change, and the resonance angle changes. The SPR measurement device measures the interaction between molecules by capturing the change in the resonance angle.

  In experiments and research in the field of biochemistry, proteins, DNA, drugs and the like are used as ligands and analytes. For example, when screening for drugs, biological substances such as proteins are used as ligands, and a plurality of types of drugs serving as analytes are brought into contact with the sensor surface to examine their interaction.

  The SPR measurement device described in Patent Document 1 below employs a Kretschmann arrangement as an optical system for making light incident on a metal film. In the Kretschmann arrangement, for example, a sensor in which a metal film is formed on a glass substrate that is a transparent dielectric is used, and the glass substrate and the prism are bonded to face the light incident surface of the metal film. The prism condenses the light irradiated toward the light incident surface so as to satisfy the total reflection condition. A ligand is fixed on the sensor surface, and a flow path for flowing an analyte solution obtained by dissolving an analyte in a solvent is disposed at a position facing the sensor surface. An analyte solution is fed into this flow path, the analyte and the ligand are brought into contact with each other, and the interaction between them is measured by detecting the attenuation of the reflected light at that time.

  When performing measurement, first, a buffer is injected into the flow path and fed to the sensor surface. In this state, signal measurement is started. Thereafter, the analyte solution is injected. The buffer in the flow path is pushed out by the injected analyte solution and discharged from the discharge port. Then, after the analyte solution is retained in the flow path for a predetermined time, the buffer is injected again, and the signal measurement is terminated. Thereby, it is possible to detect signals from detection of the baseline of the signal to the binding reaction between the analyte and the ligand to desorption.

  As a method of feeding the analyte solution to the sensor surface, it is supplied using a pipette tip. The pipette tip is relatively easy to attach and detach to the inlet of the flow path, and is suitable for a case where access to a large number of sensors is performed by repeatedly attaching and detaching.

  By the way, as a solvent (diluent) for the measurement buffer and the analyte solution, for example, various buffer solutions (buffer solutions), physiological salt solutions represented by physiological saline, and pure water are used. The The type of each of these liquids, the ph value, the type of mixture, the concentration thereof, and the like are appropriately determined according to the type of ligand. For example, DMSO (dimethyl-sulfo-oxide) is contained in physiological saline in order to make the analyte easily soluble. This DMSO greatly affects the signal level at the time of measurement. Since the measurement buffer is used for detection of the reference level, when DMSO is contained in the analyte solvent, a measurement buffer having a DMSO concentration comparable to that DMSO concentration is used.

  Such an analyte solution is often stored for a long period (for example, one year). In such a case, a concentration difference may occur between the initial DMSO concentration and the DMSO concentration at the time of measurement due to a change with time. When strict measurement is required, such a concentration difference is estimated from the signal level of the reference signal at the time of measurement when the analyte solution is injected, and the measurement data is corrected (DMSO concentration correction). .

Since the correction amount is estimated from the signal level at the time of measurement, it is not accurate. Further, when the numerical value of the correction amount is large, many errors are included. Therefore, a titer plate is used in which the opening of each well of the well plate for storing the analyte solution is covered with an evaporation preventing film so that such correction is not necessary.
JP-A-6-167443

  As described above, if the opening of each well is closed with an evaporation preventing film, an operation for removing the evaporation preventing film is required at the time of measurement, which is troublesome. Therefore, it is desired to use the film while covering the evaporation preventing film. In this case, before the analyte solution is sucked, a punching device for breaking the evaporation preventing film is required. As such a drilling tool, since the opening of each well is fine, the punching tool is moved with high accuracy by a moving mechanism such as a robot hand or an XYZ coordinate robot, and a hole is formed in the film covering the well opening. An expensive device that opens the door is required.

  It is also conceivable that the anti-evaporation film is broken at the tip of the pipette tip when inserted into the opening of each well. However, since the pipette tip is attached to the tip of the nozzle in a replaceable manner, the attachment strength is weak. For this reason, when contacting the evaporation preventing film, the pipette tip is bent with respect to the nozzle, and there is a possibility that the insertion into the well cannot be performed accurately.

  An object of the present invention is to provide a hole punch for breaking a film covering the opening of each well at a low cost.

  In the punching device of the present invention, an adapter having a mounting portion at the rear end that can be removably attached to the tip of the nozzle of the dispensing head, similar to the pipette tip that is removably attached to the dispensing head of the dispensing device. And a pointed head that is fixed to the front end of the adapter and breaks through the film. It is preferable that the adapter has a cylindrical shape and is fixed to the dispensing head by press-fitting the tip of the nozzle from the rear end of the attachment portion. The adapter is preferably a processed part obtained by cutting the tip of the pipette tip.

  It is preferable to provide a screwing portion that is inserted into the opening at the front end of the adapter and fixed to the adapter by screwing at the rear end of the pointed head. As the screwing portion, a female screw is formed on the inner peripheral surface, and it is preferable to use a nut member that is screwed into a bolt inserted from the opening at the rear end of the adapter.

  When the pipette tip is attached, the dispensing head is moved by the amount that the tip of the pipette tip is inserted into the opening of the well, and when the punch is attached, the tip of the tip breaks the film that covers the opening. The movement is controlled by each movement amount. Therefore, it is necessary to determine in advance the length from the rear end to the front end as a punching tool. Therefore, it is necessary to manage the length so that the length from the rear end to the front end is constant even with a punch made by processing a pipette tip. Therefore, a flange portion may be formed at the pointed head, and the front end of the adapter may be brought into contact with the flange portion to restrict the insertion amount of the screwing portion into the adapter.

  As the adapter, the tip may be processed into a shape that can fix the pointed head with a bolt or a screw. Further, it is preferable to use the shape obtained by cutting the tip of the pipette tip as it is because of low cost. Since the adapter uses a pipette tip, the adapter is tapered from the rear end to the front end of the cylindrical inner wall. Therefore, when a bolt is inserted from the rear end of the cylinder and the bolt is screwed to the screw fixing portion, the head portion of the bolt is pressed against the inner wall of the adapter. Thereby, the pointed head can be fixed to the adapter.

  As the tip shape of the pointed head, it is desirable to have a needle, scissors, scissors shape or the like that can be pierced and broken from above. By the way, in the needle shape, there is a possibility that the tip of the tapered pipette tip cannot be inserted only by making a fine hole in the film. Therefore, a polymorphic cross-sectional shape having at least one ridge line along the axial direction on the outer peripheral surface of the pointed head breaks the film so that the tip of the pipette tip can be inserted. This is desirable. Furthermore, it is preferable to form the tip shape of the pointed head in a square shape.

  In the present invention, similarly to a pipette tip that is detachably attached to a dispensing head of a dispensing device, an adapter having a mounting portion that can be detachably attached to the tip of a nozzle of the dispensing head, and a front end of the adapter Since it has a pointed head that is fixed to the film and breaks through the film, it can be used by being attached to a conventional dispensing device. For this reason, it can be produced at low cost and the film can be reliably broken. Further, in the invention in which the tip is fixed to the pipette tip, this can be achieved at a lower cost.

  As shown in FIG. 1, the measurement method using SPR is roughly divided into three steps: a fixing step, a measurement processing step (data reading step), and a data analysis step. The SPR measuring device includes a fixing machine 10 that performs a fixing process, a measuring machine 11 that performs a measuring process, and a data analyzer 91 (see FIG. 4) that analyzes data obtained by the measuring machine 11.

  The measurement is performed using the sensor unit 12 that is an SPR sensor. The sensor unit 12 is disposed so as to face the sensor surface 13a, the metal film 13 serving as the sensor surface 13a where the SPR is generated, the prism 14 bonded to the light incident surface 13b on the back surface of the sensor surface 13a, and the sensor surface 13a. And a flow path 16 through which the analyte is fed.

  As the metal film 13, for example, gold is used, and the film thickness is, for example, 500 angstroms. This film thickness is appropriately selected according to the material of the metal film, the emission wavelength of the irradiated light, and the like. The prism 14 condenses the light irradiated so as to satisfy the total reflection condition toward the light incident surface 13b. The channel 16 is a liquid feeding pipe bent in a substantially U shape, and has an inlet 16a for injecting liquid and an outlet 16b for discharging it. The tube diameter of the channel 16 is, for example, about 1 mm, and the interval between the inlet 16a and the outlet 16b is, for example, about 10 mm.

  The bottom of the channel 16 is open, and this open part is covered and sealed with the sensor surface 13a. A sensor cell 17 is constituted by the flow path 16 and the sensor surface 13a. As will be described later, the sensor unit 12 includes a plurality of such sensor cells 17 (see FIG. 2).

  The fixing step is a step of fixing the ligand to the sensor surface 13a. The fixing process is performed by setting the sensor unit 12 to the fixing machine 10. The fixing machine 10 is provided with a pipette pair 19 including a pair of pipette tips 19a and 19b. In the pipette pair 19, pipette tips 19a and 19b are inserted into the inlet 16a and the outlet 16b, respectively. Each of the pipette tips 19a and 19b has a function of injecting liquid into the flow channel 16 and sucking out from the flow channel 16, and when one of them performs the injection operation, the other performs the suction operation. And so on. Using this pipette pair 19, a ligand solution 21 in which a ligand is dissolved in a solvent is injected from the injection port 16a.

  A linker film 22 that binds to the ligand is formed at substantially the center of the sensor surface 13a. The linker film 22 is formed in advance at the manufacturing stage of the sensor unit 12. Since the linker film 22 serves as a fixing group for fixing the ligand, it is appropriately selected according to the type of ligand to be fixed.

  Before performing the ligand immobilization treatment for injecting the ligand solution 21, as a pretreatment, first, a buffer solution for fixing is sent to the linker film 22 to wet the linker film 22, and then the linker film 22 is moved to the linker film 22. In order to facilitate the binding of the ligand, the linker film 22 is activated. For example, in the amine coupling method, carboxymethyl dextran is used as the linker film 22, and the amino group in the ligand is directly covalently bonded to the dextran. As the activation liquid in this case, a mixed liquid of N ′-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS) is used. After this activation process, the flow path 16 is washed by the fixing buffer.

  As the solvent (diluent) for the fixing buffer and the ligand solution 21, for example, various buffer solutions (buffer solutions), physiological salt solutions represented by physiological saline, and pure water are used. The type of each of these liquids, the ph value, the type of mixture, the concentration thereof, and the like are appropriately determined according to the type of ligand. For example, when a biological substance is used as a ligand, a physiological saline in which ph is adjusted to near neutral is often used. However, in the amine coupling method, the linker film 22 is negatively (negatively) charged by carboxymethyl dextran. Therefore, the protein is positively (positively) charged so as to be easily bonded to the linker film 22. In some cases, a phosphate buffer solution (PBS: phosphatic-buffered, saline) having a strong buffering action containing a high concentration of phosphate may be used.

  After such activation processing and cleaning, the ligand solution 21 is injected into the sensor cell 17 and the ligand immobilization processing is performed. When the ligand solution 21 is injected into the flow path 16, the ligand 21 a diffusing in the solution gradually approaches the linker film 22 and binds. In this way, the ligand 21a is fixed to the sensor surface 13a. The immobilization usually takes about one hour, and during this time, the sensor unit 12 is stored in a state where environmental conditions including temperature are set to predetermined conditions. The ligand solution 21 in the channel 16 may be allowed to stand while the immobilization proceeds, but the ligand solution 21 in the channel 16 is preferably stirred and flowed. By doing so, the binding between the ligand and the linker film 22 is promoted, and the amount of the ligand immobilized can be increased.

  When the immobilization of the ligand 21a on the sensor surface 13a is completed, the ligand solution 21 is discharged from the flow path 16. The ligand solution 21 is sucked and discharged by the pipette tip 19b. After the immobilization, the sensor surface 13a is subjected to a cleaning process by injecting a cleaning liquid into the channel 16. After this washing, if necessary, a blocking liquid is injected into the flow path 16 to perform a blocking process for inactivating the reactive group to which no ligand is bound in the linker film 22. As the blocking liquid, for example, ethanolamine-hydrochloride is used. After this blocking process, the channel 16 is washed again. Thereafter, as will be described later, an anti-drying liquid is injected into the channel 16. Thereby, the sensor unit 12 is stored until measurement in a state in which the sensor surface 13a is prevented from being dried.

  The measurement process is performed with the sensor unit 12 set on the measuring machine 11. The measuring device 11 is provided with a pipette tip 26, and using this pipette tip 26, various liquids are discharged from the injection port 16 a into the flow channel 16 and injected into the flow channel 16.

  In the measurement (data reading) step, first, a measurement buffer is injected into the flow path 16. Thereafter, a predetermined amount of an analyte solution 27 in which the analyte is dissolved in a solvent is injected. The measurement buffer in the channel 16 is pushed out by the analyte solution 27 and discharged. The analyte solution 27 is allowed to stay in the channel 16 for a predetermined time, and then the measurement buffer is injected again. The drainage liquid (used analyte solution and measurement buffer) discharged from the discharge port 16b is sucked and collected by the suction pipe 30. Note that the flow path 16 may be once cleaned before the measurement buffer is first injected.

  Data reading is started immediately after the measurement buffer is first injected in order to detect a reference signal level, and is performed after the analyte solution 27 is injected and until the measurement buffer is injected again. . As a result, it is possible to measure the SPR signal from detection of the reference level (baseline), reaction state of the analyte and ligand (binding state), and desorption of the combined analyte and ligand by injection of the measurement buffer.

  As the buffer for measurement and the solvent (diluent) of the analyte solution 27, for example, various buffer solutions (buffer solutions), physiological salt solutions represented by physiological saline, and pure water are used. The The type of each of these liquids, the ph value, the type of mixture, the concentration thereof, and the like are appropriately determined according to the type of ligand. For example, DMSO (dimethyl-sulfo-oxide) may be included in physiological saline in order to facilitate the dissolution of the analyte. This DMSO greatly affects the signal level. As described above, the measurement buffer is used for detection of the reference level. Therefore, when DMSO is contained in the analyte solvent, it is preferable to use a measurement buffer having a DMSO concentration comparable to that DMSO concentration. .

  The analyte solution 27 is often stored for a long period of time (for example, one year). In such a case, a concentration difference occurs between the initial DMSO concentration and the DMSO concentration at the time of measurement due to a change over time. May end up. Therefore, in the well plate 88 for storing the analyte solution 27 in each well 88a, the opening of each well 88a is closed with the evaporation preventing film 15 (see FIG. 2). When performing a more rigorous measurement, the difference between the initial DMSO concentration and the DMSO concentration at the time of measurement is estimated from the signal level of the reference signal when the analyte solution 27 is injected, and the measurement data is corrected. (DMSO concentration correction) may be performed.

  Here, the reference signal (ref signal) is an SPR signal corresponding to a reference region provided on the sensor surface where the ligand is not fixed, and the measurement signal (act) of the measurement region that causes the reaction with the analyte when the ligand is fixed. Signal) and a reference signal. At the time of measurement, two signals of the measurement signal and the reference signal are detected, and at the time of data analysis, for example, a difference between the two SPR signals is taken and analyzed as measurement data. By doing so, for example, noise caused by disturbances such as individual differences between a plurality of sensor cells and liquid temperature changes can be canceled, and a signal with a good S / N ratio can be obtained. .

  The correction data for correcting the DMSO concentration is obtained by injecting a plurality of types of measurement buffers having different DMSO concentrations into the sensor cell 17 before injecting the analyte solution 27, and the ref signal corresponding to the DMSO concentration change at this time. And the amount of change in the level of the act signal.

  The measurement unit 31 includes an illuminator 32 and a detector 33. As described above, since the reaction state between the ligand and the analyte appears as a change in the resonance angle (incident angle of the light irradiated to the light incident surface), the illuminator 32 has various incidents that satisfy the total reflection condition. The light of the corner is irradiated to the light incident surface 13b. The illuminator 32 includes, for example, a light source 34 and an optical system 36 including a condenser lens, a diffuser plate, and a polarizing plate. The arrangement position and the installation angle satisfy the total reflection condition with respect to the incident angle of the illumination light. To be adjusted.

  As the light source 34, for example, a light emitting element such as an LED (Light Emitting Diode), an LD (Laser Diode), or an SLD (Super Luminescent Diode) is used. One such light emitting element is used, and light is emitted from this single light source toward one sensor cell. In the case where a plurality of sensor cells are measured simultaneously, a plurality of light emitting elements are arranged and used so that one light emitting element is assigned to each sensor cell. The diffusion plate diffuses light from the light source 34 and suppresses unevenness in the amount of light in the light emitting surface. The polarizing plate allows only p-polarized light that causes SPR to pass through. In addition, when using LD, when the direction of polarization of the light itself emitted from the light source is uniform, the polarizing plate is unnecessary. In addition, even when a light source with uniform polarization is used, if the direction of polarization becomes uneven by passing through the diffusion plate, the direction of polarization is aligned using a polarizing plate. The light thus diffused and polarized is condensed by the condenser lens and irradiated onto the prism 14. As a result, it is possible to cause the light incident surface 13b to enter the light rays having various incident angles without variation in light intensity.

  The detector 33 receives the light reflected by the light incident surface 13b and detects the light intensity. Since light rays are incident on the light incident surface 13b at various angles, the light rays are reflected on the light incident surface 13b at various reflection angles according to respective incident angles. When the resonance angle changes according to the reaction state between the analyte and the ligand, the reflection angle at which the light intensity attenuates also changes. For example, a CCD area sensor is used as the detector 33, and the change in the reflection angle is captured as the transition of the attenuation position of the reflected light in the light receiving surface. The detector 33 outputs the measurement data representing the reaction status thus obtained to the data analyzer 91. In the data analysis process, the measurement data obtained by the measuring instrument 11 is analyzed to analyze the characteristics of the analyte.

  For the sake of convenience, the direction of the incident light beam on the light incident surface 13b and the reflected light beam reflected there are parallel to the flow direction of the liquid in the flow channel 16 so that the configuration of the measurement unit 31 is clear. In this embodiment, the direction of the incident light beam and the reflected light beam is irradiated in a direction orthogonal to the flow direction. An illuminator 32 and a detector 33 are arranged (see FIG. 5). Of course, the measurement unit 31 may be arranged and measured as shown in FIG.

  FIG. 3 is an exploded perspective view of the sensor unit 12. The sensor unit 12 includes a flow path member 41 in which the flow path 16 is formed, a prism 14 in which the metal film 13 is formed on the upper surface, and the flow path member 41 in a state where the bottom surface is joined to the upper surface of the prism 14. The holding member 42 to be held and the lid member 43 disposed above the holding member 42.

  In the flow path member 41, for example, three flow paths 16 are formed. The flow path member 41 has a long shape, and the three flow paths 16 are arranged along the longitudinal direction thereof. This flow path 16 constitutes a sensor cell 17 (see FIG. 1) together with the metal film 13 bonded to the bottom surface thereof. Therefore, the flow path member 41 is formed of an elastic member, for example, rubber or PDMS (polydimethylsiloxane) in order to improve the adhesion with the metal film 13. Thus, when the bottom surface of the flow path member 41 is pressed against the top surface of the prism 14, the flow path member 41 is elastically deformed and the gap between the joint surfaces with the metal film 13 is filled, and the open bottom portion of each flow path 16 is filled. Is covered water-tightly by the upper surface of the prism 14. In this example, the example in which the number of the flow paths 16 is three has been described. Of course, the number of the flow paths 16 is not limited to three, and may be one or two, or four or more. But you can.

  A metal film 13 is formed on the upper surface of the prism 14 by vapor deposition. The metal film 13 is formed in a strip shape so as to face the plurality of channels 16 formed in the channel member 41. Furthermore, a linker film 22 is formed on the upper surface (sensor surface 13 a) of the metal film 13 at a site corresponding to each flow path 16. Engaging claws 14 a that engage with the engaging portions 42 a of the holding member 42 are provided on both side surfaces of the prism 14 in the longitudinal direction. By these engagements, the flow path member 41 is sandwiched between the holding member 42 and the prism 14, and is held in a state where the bottom surface and the top surface of the prism 14 are in pressure contact with each other. Thus, the flow path member 41, the metal film 13, and the prism 14 are integrated.

  In addition, protrusions 14 b are provided at both ends in the short side direction of the prism 14. The protrusion 14b is for positioning the sensor unit 12 at a predetermined storage position in the holder when the sensor unit 12 is stored in the holder.

  In the upper part of the holding member 42, a receiving port 42 b into which the tip of the pipette tip enters is formed at a position corresponding to the inlet 16 a of each channel 16. The receiving port 42b has a funnel shape so that the liquid discharged from the pipette tip is guided to each injection port 16a. Further, at a position corresponding to the discharge port 16 b of each flow channel 16, a liquid reservoir 42 d is formed that can retain the drained liquid that passes through the flow channel 16 and is discharged from the discharge port 16 b. The liquid reservoir 42d prevents the liquid from being scattered around the sensor unit 12 by temporarily retaining the liquid from the discharge port 16b. The pipette tip 19b is inserted into the liquid reservoir 42d when fixed, and the suction tube 30 is inserted when measuring. The liquid discharged to the liquid reservoir 42d is sucked and collected by these.

  In the fixing step, the ligand solution may be retained in the liquid reservoir 42d, and may be fed back to the sensor surface 13a by flowing back to the flow path 16. If it carries out like this, the fluidity | liquidity of the ligand solution in the flow path 16 will increase, and fixing efficiency can be improved.

  When the holding member 42 sandwiches the channel member 41 and engages with the prism 14, the lower surface of the receiving port 42b is joined to the inlet 16a and connected to the inlet 16a of the channel 16, while the liquid reservoir 42d is It joins with the discharge port 16b, and is connected with the discharge port 16b.

  In addition, cylindrical bosses 42c are provided on both sides of each receiving port 42b and each liquid reservoir 42d. These bosses 42 c are for fitting the holes 43 a formed in the lid member 43 to position the lid member 43. The lid member 43 is affixed to the upper surface of the holding member 42 by a double-sided tape 44 having a hole in a position corresponding to the receiving port 42b and the boss 42c.

  The lid member 43 covers the receiving port 42b communicating with the flow path 16 and the liquid reservoir 42d, thereby preventing the liquid in the flow path 16 from evaporating. The lid member 43 is formed of an elastic member such as rubber or plastic, and a cross-shaped slit 43b is formed at a position corresponding to the receiving port 42b and the liquid reservoir 42d. Since the lid member 43 is for preventing the liquid in the flow path 16 from evaporating, it is necessary to cover the receiving port 42b and the liquid reservoir 42d. The tube cannot be inserted. Therefore, by forming the slit 43b, the pipette tip and the suction tube can be inserted, and the receiving port 42b and the liquid reservoir 42d are blocked when the pipette tip and the suction tube are not inserted. ing. When the pipette tip or the suction tube is pushed into the slit 43b, the periphery of the slit 43b is elastically deformed, and the opening of the slit 43b is wide open to receive the pipette tip or the suction tube. When the pipette tip and the suction tube are removed, the slit 43b is restored to the initial state by the elastic force and closes the receiving port 42b and the liquid reservoir 42d.

  As shown in FIG. 4, in the fixing machine 10, a mounting space 51 for mounting a plurality of sensor units 12 is secured on a housing base 50 that is a base of the housing. The sensor unit 12 is subjected to all the fixing processes while being placed in the placement space 51. Accordingly, the mounting space 51 serves as a fixed stage that performs a fixing process on the sensor unit 12.

  The sensor unit 12 is set in the fixing machine 10 while being accommodated in the holder 52. The holder 52 can accommodate a plurality of (for example, eight) sensor units 12. The holder 52 is provided with a fitting portion that fits the protrusion 14 b of the sensor unit 12 and positions the sensor unit 12. Further, the bottom of the holder 52 is an opening except for support portions that support both ends of the sensor unit 12. As will be described later, in the measurement process, when the sensor unit 12 is taken out from the holder 52, a push-up member 81 (see FIG. 5) is inserted from the opening and the sensor unit 12 is pushed up.

  In the mounting space 51, for example, ten holders 52 can be arranged side by side, and pedestals 53 corresponding to the number of the holders 52 are provided. On each pedestal 53, a positioning boss for positioning the holder 52 is provided.

  The fixing machine 10 is provided with a pipette head 54 in which three pairs of pipettes 19 are connected to the head body. The pipette head 54 accesses the sensor units 12 arranged in the placement space 51 to inject and discharge liquid. Since the pipette head 54 is provided with three pipette pairs 19 each consisting of two pipette tips, liquid is simultaneously injected (and discharged) into the three sensor cells 17 included in one sensor unit 12. can do. The fixing device 10 is provided with a controller (not shown), and the controller controls the timing, suction amount and discharge amount of each pipette pair 19 for each pipette head 54. The

  The housing base 50 is provided with a head moving mechanism 56 that moves the pipette head 54 in three directions of X, Y, and Z. The head moving mechanism 56 is a known moving mechanism including, for example, a conveyance belt, a pulley, a carriage, and a motor. The head moving mechanism 56 moves up and down the pipette head 54, and moves the pipette head 54 in the Y direction together with the lifting mechanism. A Y-direction moving mechanism including a guide rail 58 that is freely held, and an X-direction moving mechanism that holds the guide rail 58 at both ends and moves the pipette head 54 in the X direction for each guide rail 58. The head moving mechanism 56 is controlled by a controller, and the controller drives the head moving mechanism 56 to control the vertical and horizontal positions of the pipette head 54.

  A plurality of liquid storage units 61 for storing various liquids (ligand solution, cleaning liquid, fixing buffer, anti-drying liquid, activation liquid, blocking liquid, etc.) to be injected into the flow path 16 are provided on the housing base 50. It has been. The number of liquid storage units is determined according to the type of liquid used. Each liquid storage unit 61 is provided with six insertion ports arranged side by side. The number of insertion holes and the arrangement pitch are determined according to the number of pipette tips of the pipette head 54 and the arrangement pitch. When injecting liquid into the sensor cell 17, the pipette head 54 accesses each liquid storage unit 61 to suck in a desired liquid, and then moves to the placement space 51 and injects it into the sensor unit 12.

  A pipette tip storage unit 63 for storing a new pipette tip 62 is provided on the housing base 50. The pipette tips constituting the pipette pair 19 are interchangeably attached to the tip of each nozzle of the pipette head 54 and are in direct contact with the liquid, so that different liquids are not mixed for each liquid to be used. The pipette tip 62 is replaced with a new one. Among these, the pipette head 54 is provided with a release drive mechanism for releasing the pipette tip, and the release of the pipette tip is automatically performed. When exchanging pipette tips, the pipette head 54 is moved to a disposal unit (not shown), where the used pipette tips are released, and then the pipette tip storage unit 63 is accessed to access unused pipettes. The chip 62 is picked up.

  Reference numeral 64 is a well plate in which a plurality of wells having uniform openings are arranged in a matrix, and temporarily stores the liquid sucked up by the pipette tip or mixes and mixes a plurality of types of liquids. Used when adjusting liquid. The well plate 64 has the same form as a well plate 88 for storing an analyte solution described later in detail.

  When starting the fixing, the casing of the fixing machine 10 is covered with a cover (not shown), and the inside of the fixing machine 10 including the placement space 51 is shielded from the outside. The temperature in the fixing machine 10 can be adjusted by a temperature controller (not shown). After the ligand is injected into the sensor cell 17, the sensor unit 12 is stored on the mounting space 51 for a predetermined time until the ligand 21a is fixed to the sensor surface 13a. During this storage, the ligand solution 21 in the flow path 16 is stirred as necessary. The degree of progress of immobilization during this period depends on the environmental conditions (temperature) of the sensor unit 12. Therefore, the internal temperature of the fixing machine 10 is kept at a predetermined temperature by the temperature controller. The set temperature, standing time, and the like are appropriately determined according to the type of the ligand 21a.

  When the fixing is completed, a buffer (cleaning liquid) is injected into the sensor cell 17. The buffer is injected into the sensor cell 17 by inserting the pipette tip 19a sucking the buffer into the slit 43b while the sensor cell 17 is filled with the ligand solution. When the buffer is discharged from the inlet 16a to the channel, the ligand solution already injected into the channel 16 is pushed out toward the outlet 16b and discharged. Then, the buffer to be discharged is sucked by the pipette tip 19b that performs the suction operation in conjunction with the injection operation of the pipette tip 19a. Thereby, replacement (replacement) of the liquid in the sensor cell 17 is performed.

  Then, after the cleaning is completed, the anti-drying liquid for preventing the ligand 21a from being dried is injected into the sensor cell 17 by the same procedure as described above, and the already injected buffer and the newly injected dry prevention liquid are injected. Replacement with liquid is performed. After the replacement, the sensor unit 12 is delivered to the measuring instrument 11 for each holder 52 in a state where the sensor surface 13a on which the ligand 21a is fixed is immersed in the drying prevention liquid. Thereby, the ligand is not dried until the measurement is started.

  As the drying preventing liquid, for example, various buffer liquids (buffer solutions), physiological salt solutions represented by physiological saline, and pure water are used. The type of each of these liquids, the ph value, the type of mixture, the concentration thereof, and the like are appropriately determined according to the type of ligand. Since this anti-drying liquid is for preventing the drying of the ligand 21a, it is sufficient to inject an amount sufficient to immerse the ligand 21a on the sensor surface 13a, but a little more in consideration of the amount of evaporation. For example, an amount sufficient to fill the flow path 16 may be injected.

  As shown in FIG. 5, the measuring machine 11 includes a holder transport mechanism 71, a pickup mechanism 72, a dispensing device 18, and the measuring machine 11 arranged on the measurement stage 74. The holder transport mechanism 71 includes a transport belt 76, a carriage 77 attached to the transport belt 76, and a plate 78 on which the holder 52 that is attached to the carriage 77 and stores a plurality of fixed sensor units 12 is placed. Consists of. The holder transport mechanism 71 transports each sensor unit 12 in the holder 52 to the standby stage by moving the plate 78 on which the holder 52 is placed in the X direction. In the standby stage, each sensor unit 12 in the holder 52 is set in a posture in which the longitudinal direction is along the direction orthogonal to the direction (X direction) in which the holder transport mechanism 71 moves the plate 78.

  The pickup mechanism 72 pushes up the sensor unit 12 waiting on the standby stage toward the clamping position located above, and the sensor unit 12 pushed up to the clamping position by the lifting mechanism 79 from both sides to measure the measurement stage. And a transport mechanism 89 for transporting to 74. The push-up mechanism 79 includes a push-up member 81 and a push-up member drive mechanism 83. The push-up member 81 is raised and lowered in the vertical direction (Z direction) by the push-up member drive mechanism 83. As described above, the bottom of the holder 52 is an opening, and the plate 78 is also hollow at a position corresponding to the opening. The push-up member 81 passes through the plate 78, enters the opening of the holder 52, rises from below toward the bottom surface of the sensor unit 12, holds the sensor unit 12 so as not to fall, and is held from the standby stage. Push up.

  The transport mechanism 89 includes a handling head 82 and a moving mechanism that moves the handling head 82 between the clamping position and the measurement stage 74. The moving mechanism includes a nut 84, a guide rod 85, a screw shaft 86, a motor 90, a position detector, a control unit 93, and the like. The handling head 82 includes a head body 82a and a gripping mechanism 92. The gripping mechanism 92 is provided in the head main body 82 a and handles the sensor unit 12. The motor 90 rotates the screw shaft 86 forward and backward. The screw shaft 86 is arranged along the Y direction orthogonal to the X direction. The nut 84 is moved in the axial direction of the screw shaft 86 according to the lead of the screw by the rotation of the screw shaft 86. A head main body 82 a is fixed to the nut 84. The rotation of the head main body 82a is stopped by a pair of guide rods 85 arranged in parallel with the Y direction.

  Although not shown, the position detector includes a shielding plate provided on the head main body 82a, and a horseshoe-shaped photoelectric sensor provided on each of the clamping position and the measurement stage. A signal output when each photoelectric sensor detects the shielding plate is sent to the controller 93. The control unit 93 monitors each signal obtained from each photoelectric sensor to control the driving of the motor 90, and the operations of the holder transport mechanism 71, the pickup mechanism 72, the head moving mechanism 73, and the measurement unit 31 are synchronized. Thus, the measuring machine 11 is controlled in an integrated manner.

  A rotary encoder is attached to the output shaft of the motor 90, and the controller 93 can detect the movement position of the head body 82a based on the pulse signal obtained from the rotary encoder. This fine feed control is used when the head main body 82a is accurately fed from the origin position where the shielding plate is detected by the photoelectric sensor on the measurement stage to a plurality of measurement positions which will be described in detail later. Instead of using the position detecting means and the rotary encoder, a pulse motor may be used, and the rotation angle, rotation direction, rotation speed, etc. of the motor may be controlled by a pulse signal sent to the motor.

  The measuring machine 11 includes an illuminator 32 and a detector 33. The illuminator 32 and the detector 33 are configured so that the direction of the incident light beam applied to the sensor unit 12 and the reflected light beam reflected by the sensor surface 13a and the arrangement direction of the sensor cells 17, that is, the flow direction of the horizontal portion of the flow path 16 are as follows. It arrange | positions so that it may orthogonally cross.

  The dispensing device 18 includes a dispensing head 87 that holds a punching pin (a punching tool) or a pipette tip 26, which will be described in detail later, and a head moving mechanism 73 that moves the dispensing head 87. At the accessible position of the dispensing head 87, a measurement buffer suction position, an analyte solution suction position, a measurement position, a punching pin (perforating tool) replacement position, and a pipette replacement position are arranged. .

  A well plate for accommodating the measurement buffer is disposed at the measurement buffer suction position. A well plate 88 for storing the analyte solution 27 is disposed at the analyte solution suction position. The well plate 88 arranged at each position is set in a direction in which each row of a plurality of wells provided in a matrix is aligned in the X and Y directions. Each well contains a solution. For example, different wells of the analyte solution 27 are accommodated in each well of the well plate 88 disposed at the analyte solution suction position, and the evaporation prevention film 15 is covered on the whole well from above (see FIG. 2). As this film 15, for example, it is desirable to use a film in which vinyl is laminated on an aluminum foil.

  When the sensor unit 12 is transported to the measurement position, the head moving mechanism 73 carries the dispensing head 87 to the measurement buffer suction position or the analyte solution suction position, and the sensor unit 12 at the measurement position. The sensor cell 17 is accessed to inject and discharge the liquid.

  A perforated pin is prepared at the perforated pin replacement position. The dispensing head 87 is provided with a nozzle that sucks and discharges liquid, and a pipette tip 26 or a perforated pin is replaceably attached to the tip of the nozzle. A suction tube 30 that is paired with the pipette tip 26 is disposed at the measurement position. The pipette tip 26 and the suction tube 30 inject and discharge liquid one by one with respect to a specific sensor cell 17 to be measured.

  Before dispensing the analyte solution with the pipette tip 26, the dispensing device 18 accesses the dispensing head 87 to the drilling pin replacement position and removes the pipette tip 26 attached to the tip of the nozzle to make the hole. A pin is attached, and the evaporation preventing film 15 is pierced with the pierced hole pin. After that, the pipette tip 26 is attached again by accessing the piercing pin replacement position, and the pipette tip 26 is inserted into the portion of the pierced film 15. Aspirate the analyte solution.

  A well plate that accommodates the measurement buffer is provided at the measurement buffer suction position, and a pipette tip storage unit that accommodates the exchange pipette tip is provided at the pipette exchange position. After injecting and discharging the liquid, the head moving mechanism 73 moves the dispensing head 87 to the pipette replacement position and replaces the pipette tip 26 attached to the nozzle with a new one. Among these mechanisms, the release of the pipette tip 26 or the perforated pin is performed by a release drive mechanism. The release drive mechanism automatically releases the pipette tip and the piercing pin from the nozzle. This mechanism is the same as that described in the fixing machine 10.

  As described above, the sensor unit 12 has a plurality of sensor cells 17. For this reason, in the measurement stage 74, the transport mechanism 89 finely sends the sensor units 12 in the Y direction at the arrangement pitch of the sensor cells 17 from the origin position, so that each sensor cell 17 sequentially reaches the measurement position on the optical path of the illuminator 32. Positioned.

  At the time of measurement, the dispensing head 87 accesses the well plate 88, sucks the desired analyte solution 27, moves the sucked pipette tip 26 to the measurement stage 74, and moves it to the sensor cell 17 at the measurement position. Analyte solution 27 is injected. The sensor unit 12 sent out from the fixing machine 10 is stored in each sensor cell 17 until it is mounted on the measuring machine 11 and measurement is started. When the measurement buffer is injected and measurement is started, this anti-drying solution is pushed out by the measurement buffer injected by the pipette tip 26, discharged from the sensor cell 17, and sucked by the suction tube 30.

  Note that when the anti-drying liquid is discharged from the flow path 16, it is preferable to inject the measurement buffer a plurality of times. This is because the anti-drying liquid cannot be completely discharged from the flow path 16 just by injecting the measurement buffer only once, and there is a high possibility that the anti-drying liquid remains. Increasing the number of injections of the measurement buffer increases the effect of discharging the anti-drying liquid, and the residual amount of the anti-drying liquid in the flow path 16 can be reduced.

  At the time of this measurement, the measurement data read by the detector 33 is transmitted to the data analyzer 91. The data analyzer 91 analyzes the reaction state between the analyte and the ligand based on the measurement data.

  As shown in FIG. 6, the dispensing device 18 includes a dispensing head 87, a Z-direction guide mechanism 24, a Z-direction moving mechanism 25, a holding unit 28, X and Y-direction moving mechanisms 29 and 59, a suction pump 39, And it is comprised by the control part 93. FIG. The head moving mechanism 73 described above includes the Z direction guide mechanism 24, the Z direction moving mechanism 25, and the X / Y direction moving mechanisms 29 and 59.

  The dispensing head 87 is provided with a nozzle 40 for sucking liquid, and the pipette tip 26 is attached to the tip of the nozzle 40 in a replaceable manner. In the present embodiment, a perforating pin (perforating tool) 20 is attached to the nozzle 40 in place of the pipette tip 26.

  The Z-direction guide mechanism 24 guides the dispensing head 87 so as to be movable in the Z direction (vertical direction). For example, the Z-direction guide mechanism 24 includes a slider provided on the dispensing head 87 and a guide rail with which the slider engages. ing. The guide rail is disposed in parallel with the Z direction (vertical direction) and fixed to the holding portion 28.

  The Z-direction moving mechanism 25 includes, for example, a motor 45 that can control the rotation angle in accordance with a control pulse given by a servo motor or a stepping motor, a feed shaft 46 to which the drive of the motor 45 is transmitted, and a feed shaft 46 And a carrier base 47 that engages with the carrier. A dispensing head 87 is fixed to the carrier table 47. The feed shaft 46 is arranged in parallel with the guide rail described above. The Z-direction moving mechanism 25 drives the motor 45 to move the dispensing head 87 in the guide direction of the Z-direction guide mechanism 24, and a retraction position for aspirating the solution from each well and retreating from the well. The pipette tip 26 is moved to the position. At the time of punching, the punching pin 20 is moved between the punching position for breaking the film 15 and the retracted position.

  The holding unit 28 integrally holds the dispensing head 87, the Z-direction guide mechanism 24, and the Z-direction moving mechanism 25. The X / Y direction moving mechanisms 29 and 59 are transport mechanisms that move the holding unit 28 in synchronization with the X and Y directions, and include X and Y direction motors. Note that the X-direction moving mechanism 29 is simply described in order to prevent complication of the drawing.

  The suction pump 39 is built in the dispensing head 87 and is driven to suck a predetermined amount of solution from the tip of the pipette tip 26 through the nozzle 40. As shown in FIG. 7, the controller 93 individually controls the suction pump 39, the release drive mechanism 38, and the X, Y, and Z motors 45, 55, and 57 via the drivers 95 to 99. .

  As shown in FIGS. 8 and 9, the punch pin 20 includes an adapter 65 and a pointed head 66. The adapter 65 is made by cutting the tip of the pipette tip 26 so that the pointed head 66 can be fixed. The pipette tip 26 is tapered and has a cylindrical cross section. The adapter 65 includes a cap (attachment portion) 67 detachably attached to the tip of the nozzle 40 at the rear end. The cap 67 is press-fitted into the outer periphery of the tip 68 of the cylindrical nozzle 40 and has the same shape as the pipette tip 26.

  The pointed head 66 has a flange 70 protruding outward from the tip 69 of the adapter 65, a pin 75 provided on one side of the flange 70, and a screwing portion 80 provided on the other side along the insertion direction of the nozzle 40. Are formed integrally. The pin 75 is formed in a narrow bowl shape, for example, a square shape having at least one ridge line on the outer surface. The screwing portion 80 has a cylindrical shape with a smaller diameter than the flange 70, and a screw hole 102 is formed at the rear end. A bolt 94 inserted from the inside of the cap 67 of the adapter 65 is screwed into the screw hole 102.

  The pipette tip 26 is hollow in order to suck the solution. When the adapter 65 is manufactured by processing, the tip 69 of the pipette tip 26 is cut at a position where the screwing portion 80 can be inserted. When the screwing portion 80 is inserted into the cut tip 69, the tip of the adapter 65 comes into contact with the flange 70, and the protruding amount of the pointed head 66 is positioned. After that, when the bolt 94 is inserted into the adapter 65 from the cap 67 and screwed into the screw hole 102 from the inside of the adapter 65, the head of the bolt 94 is caught on the inner wall (taper taper surface) of the adapter 65, The head 66 is prevented from coming off.

  The nozzle 40 is provided with a release drive mechanism 38. As shown in FIG. 10, the release drive mechanism 38 includes an extruding unit 100 that is movable in the Z direction at a part of the nozzle 40, and a driving unit 101 that drives the extruding unit 100 in the axial direction of the nozzle 40. Has been. The drive unit 101 is controlled by the control unit 93 described above via a driver 96. After the dispensing head 87 is accessed to the release position, the control unit 93 drives the drive unit 101 so that the pushing unit 100 moves from the initial position toward the pushing position and pushes the cap 67 of the adapter 65 from the nozzle 40. The punched pin 20 that has been pushed out is held in an attached posture by a catch portion provided at the release position. Note that the same procedure as described above is performed when the pipette tip 26 is released.

  When attaching the piercing pin 20 to the tip of the nozzle 40, by lowering the dispensing head 87 with the pushing portion 100 returned to the initial position, the cap 67 of the piercing pin 20 held by the catch portion This can be done by press-fitting the tip of the nozzle 40. The pipette tip 26 is attached in the same procedure as described above.

  Hereinafter, the operation of the above configuration will be described with reference to the flowchart shown in FIG. In the fixing step, the sensor unit 12 is placed in the mounting space 51 of the fixing machine 10 in a state where the sensor unit 12 is accommodated in the holder 52. In the fixing step, first, a fixing buffer is injected into each sensor cell 17 to wet the sensor surface 13a. Next, an activation liquid is injected to activate the sensor surface 13a. After washing, the ligand solution 21 is injected into the sensor cell 17 to start immobilization. In this state, the sensor unit 12 is stored on the placement space 51 for a predetermined time. During this time, the ligand 21a in the ligand solution and the linker film 22 are bonded, and the immobilization proceeds.

  When the immobilization is completed after a predetermined time has elapsed, a blocking process is performed after washing. When the blocking process is completed, the anti-drying liquid is injected into the sensor cell 17 after cleaning. The sensor unit 12 is sent to the measuring instrument 11 with the sensor surface 13a immersed in the anti-drying liquid.

  The plurality of sensor units 12 are set on the plate 78 of the measuring instrument 11 while being accommodated in the holder 52 in alignment. When a measurement start instruction is given to the measuring instrument 11, it is confirmed whether or not the holder 52 is set on the plate 78. When set, the holder transport mechanism 71 is operated to set the sensor unit 12 on the standby stage.

  When the unmeasured sensor unit 12 is set on the standby stage, the push-up member drive mechanism 83 operates to raise the push-up member 81 to the standby stage and stop. Thereafter, the push-up member drive mechanism 83 raises the push-up member 81 again toward the clamping position located further upward. As a result, the sensor unit 12 enters between the pair of claws of the handling head 82 and is sandwiched between the pair of claws. Thereafter, the sensor unit 12 is transported to the measurement stage 74 by the transport mechanism 89 while being sandwiched by the handling head 82.

  The sensor unit 12 carried to the measurement stage 74 is moved with fine accuracy in the Y direction by controlling the rotation angle of the motor 90, and the sensor cell 17 to be measured among the plurality of sensor cells 17 is set at the measurement position. The Thereafter, the dispensing head 87 is moved to the measurement buffer suction position by driving the head moving mechanism 73, the measurement buffer suction position is accessed by the pipette tip 26, and the measurement buffer is sucked. The moving mechanism 73 moves the dispensing head 87 to the measurement position and injects the measurement buffer into the sensor cell 17. At this time, the drying prevention liquid is discharged through the suction tube 30. When the measurement buffer is injected, the measurement unit 31 operates and data reading is started. After injecting the measurement buffer, the pipette tip 26 is replaced.

  Thereafter, the hole is made in the film 15, and then the analyte solution 27 is sucked and injected into the sensor cell. This series of operations will be described in detail. First, the dispensing device 18 accesses the dispensing head 87 to the drilling pin replacement position, removes the pipette tip 26 attached to the tip of the nozzle 40, and installs the drilling pin 20, and in that state X The movement of the Y-direction moving mechanisms 29, 59 causes the perforating pin 20 to reach the upper position of the well 88a containing the predetermined analyte solution in the well plate 88 arranged at the analyte solution suction position. The dispensing head 87 is moved, and after the movement is completed, the dispensing head 87 is lowered toward the punching position by the movement of the Z-direction moving mechanism 25.

  The amount of descent at this time is the amount of movement for moving the punching pin 20 to the drilling position. By this descending, the perforating pin 20 breaks through the evaporation preventing film 15 covering the opening of the well 88a as shown in FIG. This perforated position is a position where the tip of the pointed head 66 does not contact the analyte solution stored in the well 88a.

  Thereafter, the dispensing head 87 is moved to the retracted position by the movement of the Z direction moving mechanism 25, and then the dispensing head 87 is accessed again by the movement of the X / Y direction moving mechanisms 29, 59 to the punching pin replacement position. Then, the piercing pin 20 is replaced with a pipette tip 26. In these exchanges, the release drive mechanism 38 is driven to move the pushing unit 100 to push the cap 67 from the tip 68 of the nozzle 40 to release, and the tip 68 of the nozzle 40 is pressed against the cap 67 at a position different from the release position. And attach.

  Thereafter, the dispensing head 87 is moved by the X / Y direction moving mechanisms 29 and 59 so that the pipette tip 26 reaches the upper position of the well 88 a that has broken the evaporation preventing film 15. Thereafter, the dispensing head 87 is moved to the suction position by the Z-direction moving mechanism 25. Thereby, the tip of the pipette tip 26 is immersed in the analyte solution 27 stored in the well 88a as shown in FIG. Thereafter, the suction pump 39 is operated to suck the analyte solution 27 by a predetermined amount with the pipette tip 26.

  After the analyte solution is sucked, the head moving mechanism 73 moves the dispensing head 87 to the retracted position, and after the movement is completed, this time the dispensing head 87 is moved to the measurement position. When the dispensing head 87 is moved to the measurement position, the analyte solution 27 is injected into the sensor cell 17. At this time, the measuring buffer is discharged by the suction tube 30.

  When the analyte comes into contact with the sensor surface 13a, an interaction between the analyte and the ligand occurs. After a predetermined time has elapsed, after the pipette tip 26 is replaced, the measurement buffer is injected again, and the analyte solution 27 is discharged. Thereafter, the data reading ends. The measurement data obtained in this way is sent from the detector 33 to the data analyzer 91 and analyzed. Such a measurement process is sequentially performed on the plurality of sensor cells 17.

  The sensor unit 12 for which the measurement has been completed is returned again to the clamping position and is collected in the holder 52.

  After the measurement-completed sensor unit 12 is housed in the holder 52 again, the holder transport mechanism 71 operates to move the plate 78 by one pitch and carry the second sensor unit 12 to the standby stage. Hereinafter, by repeating the above-described procedure, the sensor units 12 are sequentially conveyed to the measurement stage to perform measurement.

  As described above, by using the well plate 88 in which the opening of each well 88a for storing the analyte solution 27 is covered with the evaporation preventing film 15, the evaporation of the analyte solution can be prevented. For this reason, the DMSO concentration of the analyte solution can be kept constant over the long term, and accurate measurement can be performed even in the measurement process with improved through top.

  In the above embodiment, the evaporation prevention film is used for the well plate for storing the analyte solution. However, the present invention is not limited to this, and the measurement buffer, the drying prevention solution, the ligand solution, the washing solution, and the like are stored. An evaporation prevention film may also be used for the well plate. In this case, the dispensing device having the configuration of the present invention may be used for the fixing machine.

  Further, in each of the above embodiments, the nozzles 40 are provided in the dispensing head 87 and the pipette tip 26 and the perforated pin 20 are exchanged for use. However, the present invention is not limited to this. Alternatively, the pipette tip 26 and the perforated pin 20 may be provided side by side on the dispensing head 87 so that both perforation and suction can be performed without replacement.

  Further, in the above embodiment, one pipette tip 26 is attached to the dispensing head 87 provided with a perforated pin, and the sensor cell 17 is used with the suction pipe 30 provided separately from the pipette tip 26 at the measurement position. Injecting and discharging are performed, but the present invention is not limited to this, and the dispensing head 87 may be provided with a pair of pipette tips for performing injection and discharging. In this case, the solution may be sucked from the well plate with a pipette tip for injection. Furthermore, not only a pair of pipette tips, but also a configuration in which multiple pairs of pipette tips are provided in the dispensing head may be used.

  Furthermore, in each of the above embodiments, the dispensing head 87 is moved in the Z direction, that is, in the vertical direction. However, the present invention is not limited to this, and any direction other than the X and Y directions can be used. Good. Further, as a mechanism constituting the Z-direction guide mechanism 24, the Z-direction moving mechanism 25, the X / Y-direction moving mechanisms 29, 59, etc., any mechanism can be used as long as it is a known mechanism. But it can be adopted.

  In the above example, the measurement machine 11 having the measurement stage and the fixed machine 10 having the fixed stage are described as being provided in separate housings. However, the measurement stage and the fixed stage are provided separately. If so, they may be provided in the same housing, and the dispensing device may be used in common. Further, the present invention is not limited to a dispensing device used in a measuring machine or a fixed machine, and the present invention is adopted in any dispensing device provided with a dispensing head provided with a nozzle for sucking and discharging a liquid. be able to.

  In this embodiment, the SPR sensor that generates SPR on the sensor surface and detects the attenuation of the reflected light at that time has been described as an example. However, the present invention is not limited to the SPR sensor, and the present invention is not limited to the total reflection attenuation. It can be applied to other sensors used for the measurement used. As a sensor using total reflection attenuation, for example, a leakage mode sensor is known in addition to the SPR sensor. The leakage mode sensor is composed of a dielectric, and a thin film composed of a clad layer and an optical waveguide layer that are sequentially layered thereon. One surface of the thin film serves as a sensor surface, and the other surface receives light. It becomes a surface. When light is incident on the light incident surface so as to satisfy the total reflection condition, a part of the light passes through the cladding layer and is taken into the optical waveguide layer. When a surface acoustic wave (SAW) is generated in this optical waveguide layer, the reflected light at the light incident surface is greatly attenuated. The incident angle at which the surface acoustic wave is generated varies according to the refractive index of the medium on the sensor surface, similar to the resonance angle of SPR. By detecting the attenuation of the reflected light, the chemical reaction on the sensor surface is measured.

It is explanatory drawing of a SPR measuring method. It is sectional drawing which shows the well plate which accommodated the analyte solution. It is a disassembled perspective view which shows the structure of a sensor unit. It is a perspective view which shows the outline of a fixing machine. It is a perspective view which shows the outline of a measuring machine. It is a perspective view which shows a dispensing apparatus. It is a block diagram which shows the electrical outline of a dispensing apparatus. It is a disassembled perspective view which shows a punch pin. It is sectional drawing which shows the state which attached the punch pin to the nozzle. It is sectional drawing which shows the state which removed the piercing pin from the nozzle. It is a flowchart figure which shows the procedure of a SPR measuring method. It is explanatory drawing which shows the state which broke the evaporation prevention film with the punch pin. It is explanatory drawing which shows the state inserted in the well through the film which broke the pipette tip.

Explanation of symbols

11 Measuring Machine 12 Sensor Unit 18 Dispensing Device 20 Hole Pin 26 Pipette Tip 38 Release Drive Mechanism 65 Adapter 66 Pointed Head 87 Dispensing Head

Claims (9)

A plurality of well openings containing solutions have a dispensing head that can access a well plate covered with a film, and a pipette tip that is detachably attached to the tip of a nozzle of the dispensing head is attached to the opening. Used in a dispensing device that inserts and sucks the liquid,
In the drilling tool for forming an insertion hole for inserting the tip of the pipette tip through the film before sucking the liquid,
Similar to the pipette tip, the hole comprises an adapter having a mounting portion detachably attached to the tip of the nozzle, and a pointed head fixed to the front end of the adapter and breaking through the film. Opening tool.   2. The hole according to claim 1, wherein the adapter has a cylindrical shape, and is fixed to the dispensing head by press-fitting and fitting the tip of the nozzle from the rear end of the attachment portion. Opening tool.   The punching tool according to claim 1 or 2, wherein the adapter is a processed part obtained by cutting a tip end of the pipette tip.   The screw-fixing portion that is inserted into the opening at the front end of the adapter and that fixes the peak-head to the adapter by screwing is provided at the rear end of the apex. The drilling tool according to either 2 or 3.   5. The screw fixing portion is a nut member in which a female screw is formed at a rear end in an insertion direction of the adapter and is screwed with a bolt inserted from an opening at the rear end of the adapter. Drilling tool.   The flange according to any one of claims 1 to 5, wherein a flange portion is formed on the pointed head so as to be in contact with a front end of the adapter and restrict an insertion amount of the screwing portion into the adapter. Drilling tool.   The adapter is tapered from a rear end to a front end of a cylindrical inner wall, and the pointed head is fixed by pressing the head of the bolt against the inner wall of the adapter. 6. Drilling tool according to 6.   The drilling tool according to any one of claims 1 to 7, wherein at least one ridge line is formed along the axial direction on the outer peripheral surface of the tip of the pointed head. The drilling tool according to claim 8, wherein the tip of the pointed head has a rectangular shape.

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