JP2014178252A - Inspection package, detection method and biomolecule array screening method - Google Patents

Abstract

PROBLEM TO BE SOLVED: To detect a specific reaction between a target and a biomolecule with high accuracy.SOLUTION: An inspection package includes a substrate that has a holding container for storing a sample comprising a target, and a support part that is provided in the space of the holding container, and has a biomolecule support area for supporting a biomolecule capable of specifically reacting with the target. The biomolecule support area is provided at a position lower than the upper end of a side wall forming the holding container space.

Description

  The present invention relates to a test package, a detection method, and a biomolecule array screening method.

  For example, as a well plate used for fluorescence measurement of biomolecules, a plate in which a plurality of wells are partitioned by providing a lattice-like frame member via a sealing material on a plate-like plate is known (for example, See Patent Document 1 below).

JP 2005-513457 A

However, the following problems exist in the conventional technology as described above.
When fluorescence detection is performed on a minute sample with high accuracy, it is preferable to bring the detector and the sample as close as possible. Conventionally, however, there is a limit to high-accuracy detection because the sample is arranged at the bottom of a well (eg, a recess). Therefore, it is conceivable to reduce the depth of the well. However, in this case, the amount of the sample that can be dispensed in the well decreases, and there is a possibility that the amount of the sample necessary for the reaction cannot be secured sufficiently.

  The present invention has been made in view of the above points, and provides a test package, a detection method, and a biomolecule array screening method capable of detecting a specific reaction between a target and a biomolecule with high accuracy. For the purpose.

  According to the first aspect of the present invention, a substrate having a holding container for storing a specimen containing a target, and a biomolecule support that is provided in the space of the holding container and supports a biomolecule that can specifically react with the target. And a support part having a region, wherein the biomolecule support region is provided at a position lower than the upper end portion of the side wall forming the space of the holding container.

  According to the second aspect of the present invention, the groove portion provided to be opened in the first surface is provided with a substrate in which a holding space for accommodating the specimen containing the target is surrounded by the inner surface, and the groove portion includes the target. A test package comprising: a support region that supports a biomolecule that can specifically react with the second surface at a predetermined position lower than the first surface; and a second groove portion that is open on the inner surface and communicates with the holding space. Provided.

According to the third aspect of the present invention, after dispensing the specimen into the holding space in the test package of the first aspect, the target is irradiated with light using a predetermined optical system, and the target and the biomolecule Detecting an affinity is provided.
According to the fourth aspect of the present invention, after dispensing the specimen into the holding space in the test package of the second aspect, the target is irradiated with light using a predetermined optical system to Detecting the affinity, dispensing the specimen into the holding space in the test package of the first aspect provided with the detection method, and after dispensing the specimen, the target is obtained using a predetermined optical system. And inspecting by irradiating with light.

According to the fifth aspect of the present invention, the specimen is dispensed into the holding container in the test package of the first aspect, and the affinity between the target and the biomolecule contained in the specimen after the specimen is dispensed And a method for screening a biomolecule array.
According to the sixth aspect of the present invention, the sample is dispensed into the holding space in the test package of the second aspect, and the affinity between the target contained in the sample and the biomolecule after the sample is dispensed And a method for screening a biomolecule array.

  In the present invention, a specific reaction between a target and a biomolecule can be detected with high accuracy.

FIG. 3 is an external perspective view of a chip plate CP according to the first embodiment. Sectional drawing of the well part WL which concerns on 1st Embodiment. Sectional drawing of the well part WL which concerns on 1st Embodiment. The figure which shows an example of the measuring apparatus 20. FIG. Sectional drawing of the well part WL which concerns on 2nd Embodiment. Sectional drawing of the well part WL which concerns on 3rd Embodiment. Sectional drawing of the well part WL which concerns on 4th Embodiment. The top view of the well part WL which concerns on 4th Embodiment. Sectional drawing of the well part WL which concerns on 5th Embodiment. Sectional drawing of the well part WL which concerns on 6th Embodiment. Sectional drawing of the well part WL which concerns on 7th Embodiment. Sectional drawing of the well part WL which concerns on 8th Embodiment. Sectional drawing of the well part WL which concerns on 9th Embodiment. Sectional drawing of the well part WL which concerns on 10th Embodiment. Sectional drawing of the well part WL which concerns on 11th Embodiment. A sectional view of well part WL concerning a 12th embodiment.

  Hereinafter, embodiments of the inspection package, detection method, and screening method of the present invention will be described with reference to FIGS.

[First Embodiment]
First, a first embodiment of an inspection package, a detection method, and a screening method will be described with reference to FIGS.

FIG. 1 is an external perspective view of a chip plate CP constituting the inspection package PG.
The test package PG is used for a reaction test (screening) between a target and a biomolecule contained in a specimen, and includes a chip plate (substrate) CP as shown in FIG.

  The chip plate CP is, for example, a flat plate-like substrate having a rectangular shape in plan view, and a plurality of well portions WL arranged in a matrix at a predetermined pitch (12 rows × 8 rows in FIG. 1) on the surface (first surface) CPa. I have. The material of the chip plate CP is not particularly limited as long as it has resistance to the specimen, and for example, a glass material, a plastic, a resin material, a silicon substrate, or the like can be used.

FIG. 2 shows an example of a cross-sectional detail of the well portion WL.
As shown in FIG. 2, the well portion WL constitutes a holding container having a circular shape in plan view that accommodates a specimen K containing a target (see FIG. 3). Is provided with a holding space (space) 13.

  A support portion 60 that protrudes toward the opening 15 is provided at the center of the bottom wall 11. The support unit 60 includes a support region (biomolecule support region) 61 that supports the biochip (biomolecule array) BC on a flat top surface. The support region 61 is provided at a position lower than the upper end of the side wall 12 so that a biomolecule 72 (described later) of the supported biochip BC is located at a position lower than the surface CPa. The support region 61 has an angle formed by a line connecting the support region 61 and the upper end of the side wall 12 and an optical axis of an objective lens (observation optical system) 32 to be described later, which is larger than the NA of the objective lens 32. It is provided in the position.

  The biochip (biomolecule array) BC includes a biomolecule 72 fixed to the opening 15 side of the base material 71, and an adhesive or the like is attached to the support region 61 of the support portion 60 on the back surface side of the base material 71. It is fixed with adhesive such as double-sided tape. For example, the biochip BC has a large number of probes (spots) formed by laminating a plurality of biomolecules on a 1 to 10 mm square substrate. The biochip BC is formed, for example, by forming spots on a silicon wafer and then dicing the silicon wafer into individual pieces. The probe repeats, for example, a step of arranging a predetermined biomolecule forming material on a silicon wafer and an exposure step of selectively irradiating the biomolecule forming material with light of a predetermined wavelength through a mask. Thus, it is formed by stacking a plurality of biomolecules. The biomolecule formed in this manner is contained in the specimen and reacts specifically with the labeled target, so that the measuring apparatus 20 can increase the affinity (eg, reactivity and binding property) between the target and the biomolecule. It can be detected. Further, as an example, the biomolecule formed as described above binds to the target by specifically reacting with the fluorescently labeled target of the measurement target (eg, specimen), and the predetermined light (excitation light) ) To illuminate the bound target to generate a predetermined fluorescence.

Next, prior to describing the detection method and screening method of the present embodiment, a measurement apparatus used for testing (screening) the biochip BC supported by the chip plate CP will be described.
FIG. 4 is a diagram illustrating an example of the measurement apparatus 20. In the following description, an XYZ orthogonal coordinate system is set, and the positional relationship of each member will be described with reference to this XYZ orthogonal coordinate system. The predetermined direction in the horizontal plane is the X-axis direction, the direction orthogonal to the X-axis direction in the horizontal plane is the Y-axis direction, and the direction orthogonal to each of the X-axis direction and the Y-axis direction (that is, the vertical direction) is the Z-axis direction. To do. Further, the rotation (inclination) directions around the X axis, Y axis, and Z axis are the θX, θY, and θZ directions, respectively.

  As shown in FIG. 4, the measuring device 20 includes a measuring device main body 21 for observing a chip plate CP (spot formed on the biochip BC), a control device 22 for controlling the operation of the measuring device main body 21, and a control device. And a display device 23 connected to 22. The control device 22 includes a computer system. The display device 23 includes a flat panel display such as a liquid crystal display.

  The measuring device main body 21 supports a light source device 31, an optical system 25 including an objective lens 32, and the like, and a chip plate CP in a predetermined direction (eg, X, Y, and Z axis directions, and θX, θY, and θZ directions). A movable stage (measurement stage) 26, an eyepiece 27, and an observation camera 29 including a sensor (for example, an image sensor) 28 that can receive light via an object. For example, the sensor 28 includes a photodetector such as a PMT (photomultiplier tube) and an image sensor. In the present embodiment, the sensor 28 uses an image sensor as an example. The sensor 28 can acquire image information of an object, and includes, for example, a CCD (charge coupled device) or a CMOS sensor. The measurement device main body 21 includes a body 24, and each of the light source device 31, the optical system 25, the stage 26, the eyepiece 27, and the observation camera 29 is supported by the body 24.

  The optical system 25 includes an illumination optical system 36 that illuminates the chip plate CP (biochip BC) using light emitted from the light source device 31, and a chip plate CP (biochip BC) that is illuminated by the illumination optical system 36. An image forming optical system 33 that forms an image in the vicinity of the sensor 28 and the eyepiece 27 is provided. The sensor 28 and the eyepiece 27 are disposed on the image plane side of the imaging optical system 33.

The objective lens 32 is an infinite objective lens and can face the chip plate CP supported by the stage 26. In the present embodiment, the objective lens 32 is disposed on the + Z side (upper side) of the chip plate CP.
The light source device 31 observes excitation light of a predetermined wavelength band that generates fluorescence from the chip plate CP (eg, spot formed on the biochip BC) and the chip plate CP (eg, spot formed on the biochip BC). The illumination light having a predetermined wavelength band can be emitted.

  The illumination optical system 36 uses the light emitted from the light source device 31 to illuminate the chip plate CP with excitation light or illumination light. The illumination optical system 36 includes an objective lens 32 and an optical unit 37 that can separate excitation light, illumination light, and fluorescence. The objective lens 32 emits excitation light and illumination light for illuminating the chip plate CP. The illumination optical system 36 illuminates the chip plate CP supported by the stage 26 with excitation light and illumination light from a predetermined upper side (Z direction). Further, the illumination optical system 36 transmits the fluorescence generated on the chip plate CP (spot) and guides it to the imaging optical system 33.

  The imaging optical system 33 includes an optical element 47 that separates light from the objective lens 32 and a reflection mirror 45, and forms an image of the chip plate CP in the vicinity of the sensor 28 and the eyepiece 27. The optical element 47 includes a half mirror, transmits a part of incident light, and reflects a part thereof. The optical element 47 may be a dichroic mirror. The optical element 47 may be a total reflection mirror (for example, a quick return mirror) having a function of switching the optical path.

  The stage 26 supports the chip plate CP on the object plane side of the imaging optical system 33. The chip plate CP is supported by the stage 26 so that the surface of the biochip BC supported by the support region 61 of the support portion 60 (the surface on which the biomolecule is formed) faces the objective lens 32.

  Part of the light that has entered the optical element 47 through the objective lens 32 from the chip plate CP (for example, a spot formed on the biochip BC) is transmitted through the optical element 47 and guided to the eyepiece lens 43 to be contacted. Ejected from the eye 27. An image of the biochip BC (eg, spot) is formed in the vicinity of the eyepiece 27 by the imaging optical system 33. Thereby, the observer can confirm the image of the spot through the eyepiece unit 27.

  Further, part of the light incident on the optical element 47 through the objective lens 32 and the objective lens 46 from the chip plate CP (eg, a spot formed on the biochip BC) is sequentially reflected by the optical element 47 and the reflection mirror 45. Then, the light enters the sensor 28 of the observation camera 29. An image of the biochip BC (eg, spot) is formed on the sensor 28 by the imaging optical system 33. Thereby, the sensor 28 of the observation camera 29 can acquire image information of the biochip BC (eg, spot).

  As shown in FIG. 4, the sensor 28 of the observation camera 29 and the control device 22 are connected via a cable 48, and image information (eg, image signal) of the biochip BC (spot) acquired by the sensor 28. ) Is output to the control device 22 via the cable 48. The control device 22 displays the image information from the sensor 28 using the display device 23. The display device 23 can enlarge and display the image information of the biochip BC (eg, spot) acquired by the sensor 28.

  As shown in FIG. 4, in this embodiment, the stage 26 includes a stage surface plate (plate holder) 50 that supports the chip plate CP, and a drive device that moves the stage surface plate 50 on the base member 51. 52. The stage surface plate 50 is movable on the base member 51 in the XY plane and in the Z direction. The stage 26 (drive device 52) and the control device 22 are connected by a cable 49, and the control device 22 can move the stage surface plate 50 supporting the chip plate CP within the XY plane by using the drive device 52. It is. In the present embodiment, the above-described measuring device 20 and a later-described dispenser 40 can perform a series of processing (eg, dispensing processing, reaction processing, detection processing, etc.) on the biochip BC. It is integrally configured as a screening device.

  Next, a detection method using the biochip BC supported by the chip plate CP and a screening method for the biochip BC will be described. First, as shown in FIG. 3, the sample K is dispensed (injected) from the opening 15 into the holding space 13 with respect to each well WL using a dispenser (dispensing device) 40 having a dispensing nozzle. Is done. At this time, the amount of the sample K to be dispensed is set to an amount in which the biomolecule 72 of the biochip BC is immersed in the sample K and the sample K does not overflow from the holding space 13.

  Note that before the dispensing of the specimen K into the holding space 13 is started, for example, the openings 15 of all the well portions WL are closed with the sealing material 41 (see FIG. 3). When dispensing the specimen K, the sealing material 41 is peeled off and the opening 15 is opened. After the dispensing is completed, the opening 15 is closed again by the sealing material 41, so that foreign matter is mixed into the holding space 13. Can be suppressed. Further, for example, the tip of the dispenser 40 enters the holding space 13 through the sealing material 41 with the sealing material 41 remaining without peeling off the sealing material 41 when dispensing the specimen K. It is good also as a procedure of dispensing.

  After the sample K is dispensed into the holding space 13, the reaction process between the sample K and the biomolecule 72 is performed by allowing a predetermined time to elapse at a predetermined temperature. In this reaction process, it is preferable to vibrate the chip plate CP in order to promote the reaction between the target contained in the specimen K and the biomolecule 72. When the vibration is applied to the chip plate CP, the specimen K held in the holding space 13 flows. Further, convection occurs in the holding space 13 due to a difference in the temperature of the specimen K at the time of injection, the temperature of the chip plate CP, the ambient temperature, etc., and the specimen K flows. Due to the flow of the specimen K, the reaction between the target contained in the specimen K and the biomolecule 72 is smoothly performed.

After the above reaction process is completed, the specimen K is discharged from the chip plate CP, and a cleaning process for the biochip BC is performed.
The cleaning method may be a spraying method in which a cleaning liquid is sprayed onto the chip plate CP from a cleaning nozzle, or may be an immersion method in which the biochip BC (chip plate CP) is immersed in a storage part storing the cleaning liquid. At this time, it is preferable that a flow is applied to the cleaning liquid in the storage unit so that fresh cleaning liquid is always supplied to the biochip BC.

  After the cleaning process for the biochip BC is completed, the biochip BC is dried. In the drying process, for example, a process of blowing hot air for drying from the blower fan toward the biochip BC is performed.

After the biochip BC drying process is completed, the biochip BC is measured.
The affinity between the biomolecule on the biochip BC and the target contained in the specimen K is detected at the spot (eg, biomolecule) formed on the biochip BC using the measurement apparatus 20 shown in FIG. For example, when the target is fluorescently labeled with a fluorescent dye, the measuring device 20 measures the fluorescence of the fluorescent dye generated by the excitation light. At the time of measurement, the chip plate CP is placed on the stage 26.

  The measuring apparatus 20 positions the surface of a predetermined biochip BC at a predetermined position in the Z direction, and then inserts a chip plate CP into the first imaging region where a predetermined (predetermined number) of spots on the biochip BC can be measured. Is moved in the XY plane by the stage 26, and an image of the spot is picked up using illumination light for observation.

  Next, the measuring device 20 selects and emits illumination light from the light source device 31 to illuminate the surface of the biochip BC. The illumination light emitted from the light source device 31 is separated into reflected light and transmitted light by the optical unit 37, partially reflected and partially transmitted, and after the partially reflected illumination light passes through the objective lens 32, the biochip BC. Illuminate the surface. The illumination light reflected from the surface of the biochip BC passes through the objective lens 32 and the optical unit 37 and enters the optical element 47.

  A part of the illumination light incident on the optical element 47 is transmitted through the optical element 47, guided to the eyepiece lens 43, and emitted from the eyepiece unit 27. Thereby, an image of a spot on the biochip BC is formed in the vicinity of the eyepiece 27. Further, a part of the illumination light incident on the optical element 47 is sequentially reflected by the optical element 47 and the reflection mirror 45 and enters the sensor 28 of the observation camera 29.

  As a result, images of a plurality of spots are formed on the sensor 28 in a field of view having a size corresponding to the imaging characteristics of the sensor 28 and a predetermined magnification. The sensor 28 acquires spot image information (spot light reception information) and image information (position information) of an alignment mark (not shown) provided on the biochip BC. The control device 22 stores the image information of the spot S, and obtains and stores the arrangement (X, Y, θZ) of the spot group in the field of view from the position information of the alignment mark.

  Thereafter, the measurement device 20 switches the light emitted from the light source device 31 to, for example, excitation light in a predetermined wavelength band in order to perform fluorescence measurement. The excitation light emitted from the light source device 31 is reflected (totally reflected) by the optical unit 37, passes through the objective lens 32, and then illuminates the surface (eg, spot) of the biochip BC. Fluorescence is generated from a spot that is bound by a specific reaction with a target (target) substance contained in the specimen K among spots illuminated with excitation light. The generated fluorescence passes through the objective lens 32 and the optical unit 37 and enters the optical element 47. For example, when the affinity of a spot bound by a specific reaction is high, the detected fluorescence intensity is high.

  Then, similarly to the measurement of the spot by the illumination light, the image of the spot where the fluorescence is generated is formed in the vicinity of the eyepiece 27 and in the field of view of the sensor 28. The sensor 28 acquires image information (spot light reception information) of the spot that has generated fluorescence.

  In the spot measurement using the illumination light and the spot measurement using the excitation light, the upper end portion of the side wall 12 is the highest in the well portion WL in the optical axis direction of the objective lens 32 or in a direction parallel to the optical axis (that is, on the objective lens 32). The closest position. In the present embodiment, since the support region 61 is provided at a position lower than the upper end portion of the side wall 12 and higher than the bottom wall 11, the biomolecule 72 can be immersed in the specimen K and the support region 21. Compared with the case where is provided on the bottom wall 11, measurement using the objective lens 32 having a larger numerical aperture and high resolution is possible.

  After the measurement of the first imaging area in the biochip BC is completed, the measuring device 20 moves the chip plate CP so that the biochip BC is positioned in the second imaging area adjacent to the first imaging area. The second imaging region is set at a position where a part of the alignment mark imaged in the first imaging region is imaged in the field of view of the sensor 28. And the measuring apparatus 20 implements the measurement of the spot using illumination light, and the measurement of the spot using fluorescence similarly to the imaging process with respect to the said 1st imaging area.

  Then, after performing measurement processing of a plurality of imaging areas until measurement of all spots is completed, the control device 22 synthesizes the spot measurement results of the illumination light from the measurement results of the alignment marks in each imaging area. Then, the measurement results of the spots due to fluorescence are synthesized on the screen. By comparing the measurement results synthesized on the screen, the address on the biochip BC of the spot where the target of the specimen K and the biomolecule 72 are bound by a specific reaction can be measured.

  As described above, in this embodiment, since the support region 61 that supports the biochip BC is provided at a position lower than the side wall 12, the specimen K dispensed in the holding space 13 and the biomolecule 72 can react. In addition, since the support region 61 is the lowest part of the well part WL and is provided at a position higher than the bottom wall 11, the objective lens 32 having a higher numerical aperture (NA) and high resolution is used. The biomolecule 71 can be screened with higher accuracy. Note that the measurement apparatus 20 in the present embodiment is not limited to the optical detection method described above, and may be an electrical detection method.

[Second Embodiment]
Next, a second embodiment of the chip plate CP will be described with reference to FIG.
In this figure, the same components as those of the first embodiment shown in FIGS. 1 to 4 are denoted by the same reference numerals, and the description thereof is omitted.
The difference between the second embodiment and the first embodiment is the configuration of the well portion WL.

  As shown in FIG. 5, the support part 60 in the well part WL of the present embodiment includes an inclined surface 62 that gradually goes downward from the support region 61 in a direction along the surface CPa (for example, in the horizontal direction). ing. In other words, the inclined surface 62 is formed so as to gradually rise toward the support region 61 in the horizontal direction, for example. Further, the inclined surface 62 is configured to intersect the side wall 12 of the well portion WL.

  Therefore, in the present embodiment, when a horizontal flow occurs in the specimen K in the holding space 13 by applying vibration to the chip plate CP during the reaction process, the specimen K rises along the inclined surface 62, and the support region 61. Will reach the biochip BC supported by Therefore, in this embodiment, in addition to obtaining the same actions and effects as in the first embodiment, the target (target) and the biochip BC (biomolecule 72) included in the specimen K are more effective. It becomes possible to react.

[Third Embodiment]
Next, a third embodiment of the chip plate CP will be described with reference to FIG.
In this figure, the same reference numerals are given to the same elements as those of the second embodiment shown in FIG.
The difference between the third embodiment and the second embodiment is the configuration of the well portion WL.

As shown in FIG. 6, in the well portion WL in the present embodiment, a spiral surface 62 </ b> A that gradually rises to the support region 61 is formed on the inclined surface 62 as it proceeds in a predetermined direction around the vertical axis passing through the support region 61. .
Other configurations are the same as those of the second embodiment.

  Therefore, in this embodiment, when a horizontal flow occurs in the specimen K in the holding space 13 by applying vibration to the chip plate CP during the reaction process, the specimen K rises along the inclined surface 62 and the spiral surface 62A. The biochip BC supported by the support area 61 is reached. Therefore, in this embodiment, in addition to obtaining the same actions and effects as in the second embodiment, the target contained in the specimen K and the biochip BC (biomolecule 72) can react more effectively. Become.

[Fourth Embodiment]
Next, a fourth embodiment of the chip plate CP will be described with reference to FIGS. In these drawings, the same components as those of the first embodiment shown in FIGS. 1 to 4 are denoted by the same reference numerals, and the description thereof is omitted.
The difference between the fourth embodiment and the first embodiment is the configuration of the well portion WL.

FIG. 7 is a cross-sectional view of the well portion WL, and FIG. 8 is a plan view of the well portion WL.
In the first to third embodiments, the support portion 60 protrudes from the bottom wall 11 to the holding space 13. However, in the present embodiment, the support portion 60 protrudes from the side wall 12 to the holding space 13. .
The support portion 60 of the present embodiment includes a support piece 63 including a support region 61 of the biochip BC, and a suspension portion 64 that is suspended between the support piece 63 and the side wall 12.

  The support piece 63 is rectangular in plan view and has a larger area than the biochip BC. The suspension portion 64 is formed of a linear member extending in the horizontal direction, and is supported on both sides of the support piece 63 so that the support piece 63 is positioned at the approximate center in plan view in the holding space 13. . The height of the support region 61 is set at a position lower than the upper end portion of the side wall 12 and higher than the bottom wall 11 as in the above embodiment.

  In the well portion WL having the above-described configuration, the biochip BC supported by the support region 61 is located at a position lower in the Z direction than the upper end portion of the side wall 12, so that the biomolecule 72 is included in the sample K. Can react. In addition, since the support region 61 is higher in the Z direction than the bottom wall 11, the objective lens 32 having a larger numerical aperture (NA) can be used, and the biomolecule 71 can be detected with higher accuracy. Can do. In the present embodiment, since the specimen K can be held in the entire holding space 13 between the support portion 60 and the bottom wall 11, a sufficient amount of specimen can be secured for the reaction.

[Fifth Embodiment]
Next, a fifth embodiment of the chip plate CP will be described with reference to FIG. In this figure, the same reference numerals are given to the same elements as those of the first embodiment shown in FIGS. 1 to 4, and the description thereof is omitted.
The difference between the fifth embodiment and the first embodiment is that an impurity removing device is provided at the bottom of the well portion WL.

  As shown in FIG. 9, in this embodiment, an impurity removing device 80 is provided in the well portion WL. The impurity removing device 80 includes a hole 81, a suction path 82, a negative pressure suction device 83, and an on-off valve 84. The hole 81 is provided with one end opened to the bottom wall 11 and the other end opened to the back side of the chip plate CP. The suction path 82 has one end connected to the other end of the hole 81 and the other end connected to a negative pressure suction device (suction device) 83. The on-off valve 84 is interposed in the middle of the suction path 82, and opens and closes the suction path 82 under the control of the control device 22.

In the well portion WL configured as described above, impurities contained in the specimen K injected into the holding space 13 settle and accumulate on the bottom wall 11. In the impurity removing device 80, the on-off valve 84 is opened to open the suction passage 82, and the negative pressure suction device 83 is operated. As a result, the impurities contained in the specimen K are sucked and removed from the hole 81 opening in the bottom wall 11. When the impurity recovery operation for a predetermined time is completed, the on-off valve 84 is immediately closed to close the suction passage 82 and the outflow of the sample K other than the impurities is stopped.
As described above, in this embodiment, in addition to obtaining the same operation and effect as in the first embodiment, by removing impurities contained in the specimen K, the reaction between the biomolecule 72 and the target is further improved. It can be executed with high accuracy and can contribute to improvement of screening accuracy and detection accuracy.

[Sixth Embodiment]
Next, a sixth embodiment of the chip plate CP will be described with reference to FIG. In this figure, the same reference numerals are given to the same elements as those of the first embodiment shown in FIGS. 1 to 4, and the description thereof is omitted.
The difference between the sixth embodiment and the first embodiment is that a chip support device for supporting the biochip BC on the support region 61 is provided.

As shown in FIG. 10, in this embodiment, a chip support device 85 is provided in the well portion WL.
The chip support device 85 includes a hole 86, a suction path 87, a negative pressure suction device 88, and an on-off valve 89. The hole 86 is provided with one end opened to the support region 61 and the other end opened to the back side of the chip plate CP. The suction path 87 has one end connected to the other end side of the hole 86 and the other end connected to a negative pressure suction device (suction device) 88. The on-off valve 89 is interposed in the middle of the suction path 87, and opens and closes the suction path 87 under the control of the control device 22.

  In the well portion WL configured as described above, for example, when the biochip BC is transferred to the support region 61 by a transfer device such as a mounter, the opening / closing valve 89 is opened and the suction path 87 is opened by the control of the control device 22. At the same time, the negative pressure suction device 88 is operated. Thereby, the biochip BC is adsorbed and supported by the support region 61 by the negative pressure supplied from the hole 86 opened to the support region 61.

  As described above, in this embodiment, in addition to obtaining the same operation and effect as the first embodiment, it is not necessary to separately provide an adhesive or the like in the biochip BC or the support region 61. Efficiency can be improved. In the present embodiment, since no adhesive or the like is interposed between the biochip BC and the support region 61, the biochip BC can be arranged flat, and the focusing time during measurement of the biomolecule 72 is reduced. be able to. In this embodiment, the support of the biochip BC to the support region 61 (chip plate CP) can be easily released by closing the on-off valve 89 and releasing the negative pressure suction to the biochip BC. . Therefore, in the present embodiment, the biochip BC and the chip plate CP can be easily separated after the inspection is completed, and the sorting work at the time of disposal can be made more efficient.

  Needless to say, the chip support device 85 described in this embodiment can be applied to the other embodiments described above, but the same applies to the fourth embodiment described with reference to FIGS. 7 and 8. It is applicable to. In this case, for example, a hole opening in the support region 61 is provided in the support piece 63, a suction path connected to the hole is provided in the suspension part 64, a negative pressure suction device connected to the suction path is provided, and the An open / close valve may be provided in the middle of the suction path.

[Seventh Embodiment]
Next, a seventh embodiment of the chip plate CP will be described with reference to FIG. In this figure, the same reference numerals are given to the same elements as those of the first embodiment shown in FIGS. 1 to 4, and the description thereof is omitted.

  As shown in FIG. 11, the well portion WL of the present embodiment includes a groove portion (first groove portion) 5 having a circular shape in plan view provided with the surface (first surface) CPa as the opening portion 15, and communicates with the groove portion 5. The second groove portion 8 is provided. The groove 5 is formed so as to be surrounded by an inner surface composed of a bottom surface 11A and a side surface 12A. The bottom surface 11 </ b> A includes a support region 61 at the center of the groove 5. The position (height) of the support region 61 with respect to the surface CPa is set based on the NA of the objective lens 32 described above. For example, the groove may be concave.

  The second groove 8 is arranged in a ring shape around the support region 61 and opens to the bottom surface 11A. The second groove 8 is formed so as to be surrounded by an inner surface composed of a bottom surface 8A and side surfaces 8B and 8C. A holding space 13 is formed by the groove 5 and the second groove 8.

  In the well portion WL having the above-described configuration, the biochip BC supported by the support region 61 is provided on the bottom surface 11A lower than the surface CPa, so that the biomolecule 72 can react with the specimen K. . In addition, since the biochip BC is located at a position where the support region 61 is higher than the second groove portion 8, it is possible to use the objective lens 32 having a larger numerical aperture (NA), and the biomolecule 71 can be obtained with higher accuracy. Can be detected or screened. Further, in this embodiment, since the holding space 13 is formed by the second groove portion 8 in addition to the groove portion 5, it is possible to hold the sample also in the second groove portion 8, and a sufficient amount of sample necessary for the reaction can be obtained. Can be secured.

  In the above-described embodiment, the second groove portion 8 is configured to open to the bottom surface 11A. However, the present invention is not limited to this, and a configuration provided on the side surface 12A can provide the same operations and effects. it can.

[Eighth Embodiment]
Next, an eighth embodiment of the chip plate CP will be described with reference to FIG. In this figure, the same reference numerals are given to the same elements as those of the seventh embodiment shown in FIG. 11, and the description thereof is omitted.

As shown in FIG. 12, the well portion WL of the present embodiment is a spiral that gradually rises to the support region 61 as it proceeds in a predetermined direction around the vertical axis passing through the support region 61 on the side surface 8B that forms the second groove portion 8. A surface 62B is formed.
Other configurations are the same as those of the seventh embodiment.

  In the present embodiment, in addition to the same operations and effects as in the seventh embodiment, a horizontal flow is generated in the specimen in the holding space 13 by applying vibration to the chip plate CP during the reaction process. In this case, the specimen rises along the spiral surface 62B and reaches the biochip BC supported by the support region 61. Therefore, in this embodiment, the target contained in the specimen and the biochip BC (biomolecule 72) can be reacted more effectively.

[Ninth Embodiment]
Next, a ninth embodiment of the chip plate CP will be described with reference to FIG. In this figure, the same components as those of the seventh embodiment shown in FIG.

  As shown in FIG. 13, the well portion WL of the present embodiment is provided at the intersection of the surface CPa and the side surface 8C, and gradually expands from the bottom of the groove portion 5 (that is, the upper end portion of the side surface 8C) toward the surface CPa. A tapered surface TP is provided.

  In the chip plate CP having the above-described configuration, in addition to obtaining the same operation and effect as the seventh embodiment, the opening area on the surface CPa can be increased, so that it has a larger numerical aperture (NA) and high resolution. The objective lens 32 described above can be used, and the biomolecule 71 can be screened or detected with higher accuracy.

[Tenth embodiment]
Next, a tenth embodiment of the chip plate CP will be described with reference to FIG. In this figure, the same components as those of the seventh embodiment shown in FIG.

  As shown in FIG. 14, the well portion WL of the present embodiment is provided with a protruding wall 9 that protrudes higher than the surface CPa around the groove portion 5.

  In the chip plate CP having the above-described configuration, in addition to obtaining the same operation and effect as the seventh embodiment, even when the specimen dispensed into the holding space 13 overflows from the opening 15 to the surface CPa, the chip plate CP protrudes. It is possible to prevent contamination by mixing into other well portions WL by the wall 9P, and it is possible to stably carry out high-precision biomolecule testing.

[Eleventh embodiment]
Next, an eleventh embodiment of the chip plate CP will be described with reference to FIG. In this figure, the same components as those of the seventh embodiment shown in FIG.

  As shown in FIG. 15, the well portion WL of the present embodiment includes an inclined surface 62C that gradually goes downward as the side surface 8B that forms the second groove portion 8 moves away from the support region 61 in the direction along the bottom surface 11A. It has become. In other words, the inclined surface 62 </ b> C is formed so as to gradually rise toward the support region 61 in the horizontal direction, for example.

  In the chip plate CP having the above-described configuration, in addition to obtaining the same operations and effects as in the seventh embodiment, the sample in the holding space 13 is horizontally aligned by applying vibration to the chip plate CP during the reaction process. When a flow occurs, the specimen rises along the inclined surface 62C and reaches the biochip BC supported by the support region 61. Therefore, in this embodiment, in addition to obtaining the same action and effect as in the seventh embodiment, the target (target) included in the specimen reacts more effectively with the biochip BC (biomolecule 72). It becomes possible.

  In addition, about the inclined surface 62C demonstrated in the said 11th Embodiment, it progresses to the support area | region 61 as it progresses to the predetermined direction of the surroundings of the vertical axis line which passes along the support area | region 61 similarly to 3rd Embodiment demonstrated using FIG. It is good also as a structure which provides the rising spiral surface in the inclined surface 62C. By adopting this configuration, when a horizontal flow occurs in the specimen in the holding space 13, the specimen rises along the spiral surface, so that the target included in the specimen and the biochip BC (biomolecule 72) are further effective. Can be reactive.

[Twelfth embodiment]
Next, a twelfth embodiment of the chip plate CP will be described with reference to FIG. In this figure, the same components as those of the seventh embodiment shown in FIG.

  As shown in FIG. 16, the well portion WL of the present embodiment includes the impurity removing device 80 in the fifth embodiment described with reference to FIG. 9, and the chip support device 85 in the sixth embodiment described with reference to FIG. It has both. The hole 81 in the impurity removing device 80 opens to the bottom surface 8 </ b> A of the second groove 8. Further, the suction paths 82 and 87 of the present embodiment have a configuration in which a suction path 90 connected to a negative pressure suction apparatus 91 shared by the impurity removing apparatus 80 and the chip support apparatus 85 is branched.

  In the chip plate CP having the above-described configuration, the opening / closing valve 84 in the impurity removing device 80 is opened to open the suction passage 82 and the negative pressure suction device 91 is operated. As a result, impurities contained in the specimen are aspirated and removed from the hole 81 opening in the bottom surface 8A. In the chip plate CP configured as described above, for example, when the biochip BC is transferred to the support region 61 by a transfer device such as a mounter, the opening / closing valve 89 is opened and the suction path 87 is opened by the control of the control device 22. At the same time, the negative pressure suction device 91 is operated. Thereby, the biochip BC is adsorbed and supported by the support region 61 by the negative pressure supplied from the hole 86 opened to the support region 61.

  Thus, in this embodiment, in addition to obtaining the same operation and effect as in the seventh embodiment, by removing impurities contained in the specimen K, the reaction between the biomolecule 72 and the target is higher. In addition to being able to be executed with high accuracy, it is not necessary to separately provide an adhesive or the like in the biochip BC or the support region 61, and the inspection efficiency can be improved. In the present embodiment, since no adhesive or the like is interposed between the biochip BC and the support region 61, the biochip BC can be arranged flat, and the focusing time when measuring the biomolecule 72 is reduced. Can be reduced. In this embodiment, the support of the biochip BC to the support region 61 (chip plate CP) can be easily released by closing the on-off valve 89 and releasing the negative pressure suction to the biochip BC. . Therefore, in the present embodiment, the biochip BC and the chip plate CP can be easily separated after the inspection is completed, and the sorting work at the time of disposal can be made more efficient. Furthermore, in this embodiment, since the negative pressure suction device 91 is shared between the impurity removal device 80 chip and the support device 85, it is possible to contribute to downsizing and cost reduction of the device.

  As described above, the preferred embodiments according to the present invention have been described with reference to the accompanying drawings, but the present invention is not limited to the examples. Various shapes, combinations, and the like of the constituent members shown in the above-described examples are examples, and various modifications can be made based on design requirements and the like without departing from the gist of the present invention.

  For example, the number of well portions WL in the chip plate CP shown in the above embodiment is an example, and the present invention can be applied to a single well portion or a plurality of well portions WL.

  DESCRIPTION OF SYMBOLS 5 ... Groove part, 8 ... 2nd groove part, 8A ... Bottom face, 11 ... Bottom wall (lowest part), 12 ... Side wall, 12A ... Side surface, 13 ... Holding space (space), 21 ... Support area (biomolecule support area), 52 ... Biomolecule, 62, 62C ... Inclined surface, 62A, 62B ... Helix surface, 83 ... Negative pressure suction device (suction device), BC ... Biochip (biomolecule array), CP ... Chip plate (substrate), CPa ... Surface (first surface), K ... Sample, PG ... Test package, WL ... Well part (holding container)

Claims (29)

A substrate having a holding container for containing a specimen containing a target;
A support part having a biomolecule support region provided in the space of the holding container and supporting a biomolecule capable of specifically reacting with the target,
The biomolecule support region is provided at a position lower than the upper end portion of the side wall forming the space of the holding container.
Inspection package. The biomolecule support region is provided at a position higher than the lowest part of the holding container.
The inspection package according to claim 1.   The test package according to claim 2, wherein a portion of the holding container that is lower than the biomolecule support region is provided around the biomolecule support region.   The test package according to claim 3, wherein the portion lower than the biomolecule support region includes an inclined surface that gradually goes downward as it is separated from the biomolecule support region.   The test package according to claim 3, wherein a portion lower than the biomolecule support region includes a spiral surface that gradually rises to the biomolecule support region as it proceeds in a predetermined direction around a vertical axis passing through the biomolecule support region. . The biomolecule support region is provided at a position lower than the upper surface of the specimen in the holding container.
The inspection package according to any one of claims 1 to 5. The biomolecule support region is provided at a position where an angle formed by at least an optical axis of an optical system for observing the target and a line connecting the biomolecule support region and the upper end is larger than an NA of the optical system. Be
The inspection package according to any one of claims 1 to 6. The upper end portion is a portion closest to the optical system in the holding container in the optical axis direction of the optical system or in a direction parallel to the optical axis direction when observing the target.
The inspection package according to claim 7. The support portion has a shape protruding from the surface of the holding container into the space of the holding container.
The inspection package according to any one of claims 1 to 8.   The inspection package according to claim 9, wherein the support portion has a shape protruding from the surface of the side wall into the space.   The test package according to claim 10, wherein the support portion forms a space for accommodating the specimen between the lowest portion of the holding container.   The inspection package according to any one of claims 1 to 11, further comprising a suction device that sucks the space of the holding container under a negative pressure.   The test package according to any one of claims 1 to 12, wherein a biomolecule array having the biomolecule is supported in the biomolecule support region.   The test package according to claim 13, further comprising: a support device configured to support the biomolecule array in the biomolecule support region by sucking an opening opening in the biomolecule support region under negative pressure. In the groove provided to be opened in the first surface, a holding space for accommodating the specimen containing the target is provided, and the substrate is formed surrounded by the inner surface,
A support region that supports biomolecules that can specifically react with the target at a predetermined position lower than the first surface, and a second groove portion that is open to the inner surface and communicates with the holding space. When,
Package for inspection. The inner surface includes a bottom surface disposed at the bottom of the holding space and a side surface disposed around the holding space,
The support region is provided on the bottom surface;
The inspection package according to claim 15, wherein the second groove portion is provided around the support region on the bottom surface.   The inspection package according to claim 16, wherein the second groove portion includes an inclined surface that gradually goes downward as the second groove portion moves away from the support region in a direction along the bottom surface.   The inspection package according to claim 16, wherein the second groove portion includes a spiral surface that gradually rises to the bottom surface as it proceeds in a predetermined direction around a vertical axis passing through the support region.   19. The inspection according to claim 15, further comprising a tapered surface that is provided at an intersection between the first surface and the inner surface and gradually increases in diameter from the bottom of the groove toward the first surface. package.   The inspection package according to any one of claims 15 to 19, further comprising a projecting wall provided around the groove and projecting higher than the first surface.   The inspection package according to any one of claims 15 to 20, further comprising a suction device that sucks the second groove part under a negative pressure.   The test package according to any one of claims 15 to 21, wherein a biomolecule array having the biomolecule is supported in the support region.   23. The test package according to claim 22, further comprising a support device that sucks an opening opening in the support region under negative pressure to support the biomolecule array in the support region.   The target and the biomolecule are obtained by irradiating the target with light using a predetermined optical system after dispensing the sample into the holding container in the inspection package according to any one of claims 1 to 14. Detecting an affinity with.   25. The detection method according to claim 24, wherein a position of the biomolecule support region that is lower than the upper end portion is set based on an NA of the predetermined optical system.   24. After dispensing the specimen into the holding space in the test package according to claim 15, the target and the biomolecule are irradiated with light using a predetermined optical system. Detecting an affinity with.   The detection method according to claim 24, wherein the predetermined position lower than the first surface that supports the biomolecule is set based on an NA of the predetermined optical system. Dispensing the sample into the holding container in the test package according to any one of claims 1 to 14;
Detecting the affinity between the target and the biomolecule contained in the sample after dispensing the sample;
A method for screening a biomolecule array comprising: Dispensing the sample into the holding space in the test package according to any one of claims 15 to 23;
Detecting the affinity between the target and the biomolecule contained in the sample after dispensing the sample;
A method for screening a biomolecule array comprising:

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