WO2017169714A1 - Dispositif d'inspection, appareil d'inspection et procédé d'inspection - Google Patents

Dispositif d'inspection, appareil d'inspection et procédé d'inspection Download PDF

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Publication number
WO2017169714A1
WO2017169714A1 PCT/JP2017/010091 JP2017010091W WO2017169714A1 WO 2017169714 A1 WO2017169714 A1 WO 2017169714A1 JP 2017010091 W JP2017010091 W JP 2017010091W WO 2017169714 A1 WO2017169714 A1 WO 2017169714A1
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Prior art keywords
pores
inspection device
inspection
substance
substrate
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PCT/JP2017/010091
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English (en)
Japanese (ja)
Inventor
孝明 伊藤
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富士フイルム株式会社
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Publication of WO2017169714A1 publication Critical patent/WO2017169714A1/fr

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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64—Fluorescence; Phosphorescence
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
    • G01N21/77—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator
    • G01N21/78—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator producing a change of colour
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals

Definitions

  • the present invention relates to a test device for detecting an antigen, antibody, deoxyribonucleic acid, or the like, which is a test substance, a test apparatus equipped with the test device, and a test method.
  • the binding phenomenon between a test substance and a capture substance is optically determined.
  • a method of detecting is known. In this method, a test substance is bound to a capture substance fixed at a predetermined position and emits fluorescence upon receiving excitation light, or a label that catalyzes a substrate reaction to generate color, fluorescence, or chemiluminescence. Is applied to the test substance, and light generated due to the label is detected.
  • a method of detecting a fluorescence generated from a fluorescent label by attaching a fluorescent label to a binding substance such as an antibody that specifically binds to the test substance, a binding of an antibody that specifically binds to the test substance
  • a binding substance such as an antibody that specifically binds to the test substance
  • a binding of an antibody that specifically binds to the test substance There are known methods for detecting color development, fluorescence, chemiluminescence, etc. generated from a chromogenic substrate, fluorescent substrate, or chemiluminescent substrate that reacts with this enzyme as a catalyst. Identification becomes possible.
  • a multi-item simultaneous detection chip in which biologically related molecules are regularly arranged on a two-dimensional substrate has been conventionally used.
  • a device made of a porous substrate in which a large number of through-holes (pores) are arranged on a support has been studied.
  • a porous silicon (Si) base material As a porous substrate, a porous silicon (Si) base material is known, but silicon has low reflectivity, and the light generated in the pores is reflected multiple times in the pores. Since the signal intensity is greatly attenuated, there is a problem that the light extraction efficiency of the optical signal generated in the pore is very low.
  • Patent Document 1 As a means for solving the above problem, porous silicon is thermally oxidized to be locally transparent silicon oxide (SiO 2 ), and the oxidized region is surrounded by a frame (wall) made of silicon.
  • a configuration has been proposed in which the light output aperture is substantially widened to improve the light extraction efficiency.
  • the outermost surface material (first layer) of the pore wall is made of a material having a refractive index larger than the refractive index of the lower layer (second layer).
  • a laminated structure has been proposed. With this laminated structure, the light reflectance on the inner wall surface can be improved, and as a result, the light detection efficiency can be improved.
  • Patent Document 1 it is possible to improve the light extraction efficiency accompanying an increase in the light aperture ratio, but the light incident on the silicon wall is also attenuated because the reflectance of silicon used as the frame wall is low. Therefore, the extraction efficiency of light incident on the frame wall is not improved.
  • the aspect ratio with the opening diameter As the aspect ratio of the pores increases, the aspect ratio of the region surrounded by the silicon frame also increases accordingly, so the light exit aperture of the region surrounded by the silicon frame becomes substantially narrow, The influence of optical signal attenuation due to multiple reflection is increased.
  • Patent Document 2 discloses a specific device manufacturing method in which holes are formed by electrochemical etching in a support constituting the second layer, and the first layer is formed by CVD (Chemical Vapor Deposition). Has been.
  • CVD Chemical Vapor Deposition
  • material restrictions are large, the second layer is substantially limited to a silicon-based material, and the first layer is limited to a material that can be formed by CVD film formation. May be high.
  • the inspection device of the present invention is an inspection device having a plurality of pores penetrating from one surface of the plate-like substrate to the other surface, A cavity formed around the region extending in the depth direction of the pores, not penetrating at least the other surface, and including at least some of the plurality of pores when viewed from one surface.
  • Have The portion of the plate-like base material constituting the region including at least a part of the pores has light transmittance,
  • This is a test device in which a capture substance that specifically binds to a specific substance is fixed to the inner wall surface of at least some of the pores.
  • the plate-like substrate is preferably made of SiO 2 or Al 2 O 3 (alumina).
  • a plurality of pores are arranged in a plurality of regions partitioned by the cavity, and one of the plurality of regions It is preferable that the same kind of trapping substance as the trapping substance is bonded to the inner wall surface of each of the pores, and the plurality of regions have areas where the trapping substances are different from each other.
  • the hollow portion may be formed so as to extend from one surface of the plate-like substrate to the front of the other surface, and may be non-penetrating on the other surface.
  • the inspection device of the present invention is a member whose material is different from that of the plate-like substrate in the opening of the other surface side of the through hole in which the hollow portion is provided so as to penetrate from one surface of the plate-like substrate to the other surface. May be blocked by the other surface and non-penetrated on the other surface.
  • the capture substance is preferably an antigen, an antibody, or deoxyribonucleic acid (DNA).
  • the equivalent circle diameter of the open region on one surface of the pore is preferably 1 ⁇ m to 100 ⁇ m.
  • the thickness of the plate-like substrate is preferably 100 ⁇ m to 2000 ⁇ m.
  • the inspection apparatus of the present invention includes the inspection device of the present invention, A solution supply unit for supplying an inspection solution into the pores of the inspection device;
  • the inspection apparatus includes a photodetector that is disposed on one surface or the other surface of the inspection device and detects light emitted from the inspection device.
  • a specimen liquid containing a specific substance is supplied to the at least some of the pores of the test device of the present invention to bind the specific substance to the capture substance
  • a labeling substance that specifically binds to a specific substance is bound to the specific substance
  • the inspection method detects light emitted from the inspection device in a state where the inspection solution is supplied to the pores and the inspection solution is retained in the pores.
  • a substance containing an enzyme label is used as the labeling substance, a reaction solution containing a substrate that is catalyzed by the enzyme label and reacts as the test solution, and the light emitted from the inspection device
  • the light produced when the substrate in the reaction solution is catalyzed by the enzyme label may be detected.
  • the substrate include a chromogenic substrate, a fluorescent substrate, and a chemiluminescent substrate, and these substrates are appropriately selected depending on the type of enzyme label.
  • the light emitted from the inspection device differs depending on the substrate, and the detected light is light absorption (coloration), fluorescence, or chemiluminescence.
  • a substance containing a fluorescent label is used as a labeling substance, Irradiate the test device with excitation light that excites the fluorescent label, As the light emitted from the inspection device, fluorescence generated from the labeling substance by irradiation with excitation light may be detected.
  • An inspection device of the present invention is an inspection device having a plurality of pores penetrating from one surface of a plate-like substrate to the other surface, extending in the depth direction of the pores, and non-existing on at least the other surface. It is a penetration and has a cavity formed around a region including at least some of the plurality of pores when viewed from one side, and constitutes a region including at least some of the pores Since the plate-like base material portion is light transmissive, it is possible to improve the extraction efficiency of light generated in the pores to at least one surface side of the inspection device. In addition, since the base material can be made of a single material, it can be manufactured at low cost.
  • FIG. 1 is a perspective view of an inspection device according to a first embodiment of the present invention. It is a top view of the inspection device shown in FIG. It is sectional drawing of the test
  • FIG. 1 is a perspective view of an inspection device according to a first embodiment of the present invention
  • FIG. 2 is a plan view of the inspection device of FIG. 1
  • FIG. 3 is a sectional view taken along line III-III of the inspection device of FIG. 4 is a bottom view of the inspection device of FIG.
  • the inspection device 1 of the present embodiment extends in the depth direction of the pores 14 on the plate-like substrate 10 having a plurality of pores 14 penetrating from one surface 11 to the other surface 12, and at least the other surface. 12 has a cavity 16 formed around a region including at least some of the plurality of pores 14 when viewed from one surface 11.
  • a total of nine pores 14 in the vertical and horizontal directions are used as one region 20, and the cavity 16 is provided so as to surround each region 20.
  • a trapping substance 30 that specifically binds to a specific substance is fixed to the inner wall surface 14 a of the pore 14.
  • the plan view shown in FIG. 2 is a view of the inspection device 1 as viewed from one surface 11, and the bottom view of FIG.
  • the inspection device 1 is a view of the inspection device 1 as viewed from the other surface 12.
  • the external shape of the inspection device 1 is configured by the external shape of the plate-like base material 10
  • one surface 11 and the other surface 12 of the plate-like base material 10 are hereinafter referred to as one of the inspection devices 1.
  • the surface and the other surface are hereinafter referred to as the surface and the other surface.
  • a cavity is formed around a region including at least a part of the pores. That is, in the inspection device, it is sufficient that a pore region surrounded by the cavity is formed at least in part. Further, in the region surrounded by the cavity, the plate-like base material constituting the pores and the cavity, that is, the pores and the wall constituting the cavity have light transmittance.
  • the entire plate-like substrate may be made of a light transmissive material, or only the region surrounded by the cavity may be made of a light transmissive material.
  • having light transmittance means that the transmittance of detection light described later is 50% or more. Specifically, the average transmittance in the visible light wavelength range of 400 to 700 nm is 50% or more. If there is, it will be regarded as having light transparency.
  • the plate-like base material is preferably made of SiO 2 or Al 2 O 3 . If the whole plate-shaped base material is comprised from the same material, preparation will be easy and it is especially preferable, but the plate-shaped base material may be provided with the part which consists of a different material. For example, only the region surrounded by the cavity is made of SiO 2 and the other part is made of Si, or only the region surrounded by the cavity is made of Al 2 O 3 and the other part is made of Al. A substrate or the like may be used.
  • the cavity 16 is a space that is open on one surface 11 side and closed on the other surface 12 side.
  • the hollow portion 16 is formed to extend from one surface 11 of the plate-like base material 10 to the front of the other surface 12, and an opening connected to the hollow portion 16 is provided on the other surface 12 of the plate-like base material 10. (See FIG. 4).
  • “formed to extend to the front” means that it extends toward the other surface 12 but does not reach the other surface 12.
  • the length of the cavity 16 extending from the one surface 11 to the other surface 12 side, that is, the depth along the depth direction of the pores 14 is at least half the thickness of the plate-like substrate 10. Is more preferable, and more preferably 3/4 or more.
  • the thickness of the plate-like substrate 10 is not particularly limited, but is preferably about 100 ⁇ m to 2000 ⁇ m.
  • the pores 14 provided in the plate-like substrate 10 are preferably arranged in an aligned manner as shown in the present embodiment, but may be arranged randomly.
  • in the region 20 surrounded by the cavity 16 hereinafter referred to as “inspection region 20”
  • nine pores of 3 rows ⁇ 3 columns are provided in one inspection region 20 for easy visual recognition, but, for example, 100 10 rows ⁇ 10 columns in one inspection region.
  • the inspection area 20 provided in one inspection device 1 may be singular or plural. Although four inspection areas 20 are shown in the present embodiment, for example, ten or more inspection areas can be provided.
  • the opening and cross-sectional shape of the pore 14 are square in this embodiment, but are not limited to a rectangle such as a square or a rectangle, but may be a circle, an ellipse, a triangle, or a polygon that is a pentagon or more. Note that the corners of the polygon may be rounded due to manufacturing reasons.
  • the pores 14 are generally columnar and the cross-sectional shape does not change, but the cross-sectional shape may partially change or the cross-sectional size may change.
  • the equivalent circle diameter of the opening in at least one surface 12 of the pore 14 is preferably about 1 ⁇ m to 100 ⁇ m. More preferably, it is 3 ⁇ m to 50 ⁇ m, and particularly preferably 5 ⁇ m to 30 ⁇ m.
  • the equivalent circle diameter refers to the diameter of a circle having an area equivalent to the area of the opening region.
  • the test substance (target molecule) to be tested in this testing device 1 is mainly a biological molecule, such as proteins such as antigens and antibodies, saccharides, peptides, DNA, ribonucleic acid (RNA), peptides Examples thereof include nucleic acids (peptide nucleic acid: PNA).
  • acquisition substance 30 specifically couple
  • the test device 1 is suitable for an allergy test provided with an allergen that is a kind of antigen as a capture substance.
  • a plurality of pores 14 are arranged in a plurality of inspection regions 20 partitioned by a cavity 16 as shown in a view of the inspection device 1 viewed from one surface 11.
  • a trapping substance of the same type is bound (fixed) to the pores 14 in one inspection region 20.
  • Different capture substances can be fixed between the examination regions 20. Accordingly, it is possible to simultaneously inspect a plurality of test substances with one inspection device 1. Note that there may be two or more regions containing the same type of capture substance between a plurality of inspection regions.
  • the same kind of allergen that is a capture substance is fixed to one or a plurality of examination areas 20, and a plurality of allergen examination areas are provided in one examination device. Thereby, the reaction with respect to a several allergen can be test
  • the cavity 16 is provided over the entire circumference of the inspection region 20, but the cavity 16 does not have to be provided over the entire circumference.
  • the total length S 1 of the portions facing the inspection region 20 of the opening of the cavity 16 disposed around the inspection region 20 is the inspection region 20.
  • the “cavity formed around the region” means a cavity arranged so as to satisfy S 1 ⁇ S 2 .
  • S 2 is , is preferably 1/4 or more the length S 3 of the pores outer peripheral edge, 1/2 or more preferably of S 3, and particularly preferably S 3 or more.
  • one side of the plurality of pores 14 arranged on the outermost periphery among the plurality of pores 14 in the inspection region 20 is a 0 , and the cavity portion 16 of the pore 14 arranged on the outermost periphery.
  • the total length S 2 12a 0 of the portions facing the surface.
  • a pore outer peripheral edge 22 shown by a broken line in the drawing
  • the length S 3 of the outer circumferential edge 22 of the pore is 4a.
  • A>a> 3a 0 S 1 ⁇ S 2 is satisfied.
  • FIG. 5 is a plan view of the inspection device 2 of the second embodiment in which the shape of the cavity is different from that of the inspection device 1 of the first embodiment.
  • the inspection region 20 including the plurality of pores 14 is configured by a plurality (here, four) of isolated rectangular cavities 17.
  • the inspection region 20 including the plurality of pores 14 is the same as that shown in FIG. 2, and the cavity 17 of the plurality of pores 14 arranged on the outermost periphery among the plurality of pores 14 in the inspection region 20.
  • the length of the portion of the opening of one cavity portion 17 facing the inspection region 20 is the length B of the long side of the rectangular cavity portion 17 and faces the inspection region 20 for the four cavity portions 17.
  • the opening shape of the pores 14 and the opening shape of the hollow portion 17 are not limited to a rectangle.
  • FIG. 6 shows a case where the pore 114 and the cavity 116 both have a circular opening, and the region 120 including the plurality of pores 114 is surrounded by the plurality of cavities 116.
  • S 1 , S 2 and S 3 are defined as follows.
  • the outer peripheral edge 122 of the pores is represented by a straight line in contact with the pores 114 arranged on the outermost periphery among the plurality of pores 114 and a line passing through the edges of some of the pores 114. .
  • the length of the portion of the pore 114 facing the cavity 116 disposed on the outer periphery (outermost periphery) 122 of the pore is the length of the arc facing the cavity 116 in the circumference of the opening of each pore 114. In this example, it is a semicircle or 3/4 circular arc portion indicated by a thick line in FIG.
  • the lengths of the portions of the openings of the plurality of cavities 116 arranged so as to surround the region 120 facing the inspection region 120 are the lengths of the arcs facing the inspection region 120 in the circumference of the openings of the cavities 116, respectively. In this example, it is a semicircle or a quarter of a circular arc indicated by a thick line in FIG.
  • the pores in the inspection region can be defined in the same manner as in FIG.
  • FIG. 7 is a plan view of the inspection device 3 according to the third embodiment
  • FIG. 8 is a cross-sectional view taken along line VIII-VIII of the inspection device 3 of FIG.
  • the pores 14 and the cavities 18 are alternately arranged vertically and horizontally.
  • one pore 14 is arranged in an inspection region 28 surrounded by four cavities 18.
  • the pore 14 and the cavity 18 have the same opening shape.
  • the pore 14 is a through hole, and the cavity 18 is provided as a blind hole that does not penetrate the other surface 12 of the plate-like substrate 10.
  • FIG. 9 shows a design change example.
  • the cavity 118 is formed on the plate-like substrate 10 after the through-hole 40 penetrating from one surface 11 to the other surface 12 is provided, and then on the other surface 12 side.
  • the opening may be formed so as to be closed with a closing member 42 made of a material different from that of the plate-like substrate 10. That is, in FIG. 9, the cavity 118 is constituted by the through hole 40 and the closing member 42 provided in the plate-like base material 10.
  • the cavity 118 is made of a material whose opening on the one surface 11 side of the plate-like substrate 10 is different from that of the plate-like substrate 10.
  • the second closing member 44 may be closed. That is, the cavity 118 may be a closed space.
  • the inside of the cavity may be a vacuum, or a gas such as air having a refractive index of approximately 1.0 may be enclosed.
  • the material constituting the closing member 42 and the second closing member 44 constituting the hollow portion 118 is not particularly limited, but it can be configured as a closing member by being hardened after being dropped by an ink jet device, such as a metal or a resin material. Material is preferred.
  • the inspection device including one pore in the inspection region surrounded by the cavity has been described.
  • a plurality of pores are included in the inspection region surrounded by the cavity. It is more preferable to have.
  • FIG. 11 is an enlarged schematic view of a part of the inspection device 1.
  • a test substance is bound to a capture substance (not shown in FIG. 11) fixed to the inner wall surface 14a of the pore 14, and the test substance is further labeled with an enzyme. It is used in a test method for detecting the presence or absence of a test substance by detecting the chemiluminescence using a labeling substance as a catalyst by binding the substance or detecting the presence and quantity of the test substance.
  • the detection of the optical signal from the present inspection device 1 is performed in a state where the pore 14 is filled with an inspection solution 61 such as a buffer solution. In addition, it uses so that the solution 61 may not enter into the cavity part 16, and the cavity part 16 comprises an air layer.
  • the base material forming the pores 14 in the inspection region 20 has light transmittance, the surface of the light generated in the pores 14 is compared with the pores 14 formed of the base material impermeable to silicon.
  • the exit aperture ratio increases.
  • the refractive index of the light-transmitting plate-like substrate 10 and the refractive index of the test solution are equivalent in the range of about 1.3 to 1.6, whereas the refractive index of the air layer is about 1. .0. That is, since the refractive index difference between the plate-like substrate 10 and the air layer is very large, a large reflection occurs at the interface.
  • the plate-like substrate 10 since the plate-like substrate 10 has a higher refractive index than the air layer, light incident on the interface C 1 at an incident angle ⁇ 1 greater than the critical angle from the plate-like substrate 10 side toward the air layer is totally reflected. Then, it goes to the surface side of the plate-like substrate 10. Even when the incident light is incident on the interface C 1 from the plate-like substrate 10 side at an incident angle ⁇ 2 smaller than the critical angle, a part of the incident light is specularly reflected at the interface C 1 and It will go to the surface side.
  • the light transmitted through the interface C 1 is incident on the plate-like base material 10 constituting the cavity 16 from the air layer, and at least a part of the light is regularly reflected at the interface C 2 , and the surface of the plate-like base material 10 is again formed. Head to the side.
  • a function of condensing light generated in the pores 14 toward the surface of the plate-like substrate 10 is provided.
  • the amount of light emitted from the surface of the inspection device 1 is increased, and the light extraction efficiency is improved.
  • the surface of the base material the surface of the inspection device.
  • the light amount can be reflected at the interface between the plate-like base material and the air layer with almost no attenuation, and the light can be condensed on the surface side of the device.
  • the light extraction efficiency can be greatly improved as compared with the case where a frame made of silicon is provided.
  • a plate-like substrate made of SiO 2 having a plurality of pores and cavities can be produced as follows. First, a plate-like substrate made of Si is prepared, and pores and cavities are formed from one surface side by wet etching or dry etching. For example, a microporous Si manufacturing process known in MEMS (Micro Electro Mechanical System) technology can be used. In the case where the cavity and the pore as shown in FIG. 2 or FIG. 5 are manufactured, when the pore is formed by etching, a mask is provided outside the pore formation region and the cavity is formed by etching. In this case, the mask may be formed sequentially with a mask other than the cavity forming region. The cavity can be formed by etching from one side and stopping the etching before the other side.
  • MEMS Micro Electro Mechanical System
  • the pore and the cavity may be etched at the same time. Since the etching speed of the cavity portion having a small diameter is slower than the etching speed of the pore portion, the pore portion penetrates the other surface, but the cavity portion can be left unpenetrated.
  • a large cavity can be formed by etching in the lateral direction so as to connect the holes.
  • the whole base material can be made transparent by changing the base material made of Si having pores and cavities formed as described above to SiO 2 by oxidation treatment.
  • oxidation treatment for example, thermal oxidation treatment is performed at a temperature of 1100 ° C. for a predetermined time.
  • the thermal oxidation treatment time may be appropriately adjusted according to the size and shape of the plate-like substrate and the desired oxidation treatment region.
  • the cavity 118 is constituted by the through hole 40 and the closing member 42 as shown in FIG. 9, first, the through hole 40 constituting a part of the cavity is formed by etching simultaneously with the pore 14. . Thereafter, the Si base material is changed to SiO 2 by the above thermal oxidation treatment, and then the opening on the other surface side of the through hole 40 constituting a part of the cavity is closed with a closing member 42 made of a material different from the base material. . As a result, the cavity 118 with the other surface closed can be formed.
  • the blocking member 42 only needs to be able to block the opening on the other surface side of the through hole 40, and may be formed so as to partially enter the through hole 40.
  • the closing member 42 can be formed, for example, by dripping a metal or a resin material so as to close the opening on the other surface side of the through hole using an ink jet device, and solidifying or curing.
  • a plate-like substrate made of Al 2 O 3 having a plurality of pores and cavities can be produced as follows.
  • a base material made of Al By preparing a base material made of Al and subjecting this base material to known anodization treatment and power wide treatment described in, for example, JP-A-2015-132737, JP-A-2014-198453, etc., A plate-like substrate made of porous alumina can be produced.
  • an Al base material is anodized to form a porous alumina coating.
  • a pore having a desired diameter is formed by performing a pore wide process to widen the diameter of the micropore.
  • the pores used as the cavity may be provided with a blocking member that covers the opening on the other surface side as in the case of the device made of SiO 2. .
  • the trapping substance which is a specific binding substance that specifically binds to the test substance, is fixed to the inner wall surface of the pores of the plate-like substrate having a plurality of pores and cavities produced as described above.
  • an inspection device can be manufactured.
  • a method for fixing the trapping substance to the inner wall surface of the pore a known method can be applied without particular limitation. For example, the method disclosed in Patent Document 1 or Patent Document 2 described above can be used.
  • FIG. 12 is a diagram schematically showing the configuration of the inspection apparatus 50 according to an embodiment of the present invention.
  • the inspection apparatus 50 according to the present embodiment is disposed on the inspection device 1, the solution supply unit 60 that supplies the inspection solution into the pores 14 of the inspection device 1, and the one surface 11 side of the inspection device 1.
  • a photodetector 70 a photodetector 70.
  • the light detector 70 detects light emitted from the inspection device 1 and is disposed on one surface 11 side of the inspection device 1 here, but is disposed on the other surface 12 side. It may be.
  • the inspection apparatus 50 is not limited to the inspection device 1 and may include any of the inspection devices described above.
  • FIG. 13 is a diagram illustrating a schematic configuration of the solution supply unit 60.
  • the solution supply unit 60 stores the inspection solution 61 installed on the other surface 12 side of the inspection device 1, and the inspection installed in the upper part of the storage unit 62 and stored in the storage unit 62.
  • the pipette part 64 for sucking the solution 61 for use, and the pressure of the depressurizing space part 66 and the depressurizing space part 66 arranged on the one surface 11 side of the inspection device 1 installed on the pipette part 64 are reduced.
  • a pump 68 for pressurization is provided.
  • the reduced pressure space 66 is decompressed by the pump 68, whereby the inspection solution 61 passes through the pipette part 64 and the pore 14 of the inspection device 1. Supplied in. Note that the inspection solution 61 is supplied to the pores 14 of the inspection device 1 and the inspection solution 61 is discharged from the pores 14 by depressurization and pressurization of the reduced pressure space 66 by the pump 68. it can.
  • the test solution 61 is a solution supplied into the pores 14 at the time of light detection, but the solution supply unit 60 includes a sample solution, a label solution, a cleaning solution, and the like to be supplied into the pores 14 in the test process. It is also used for supply. That is, the solution supply unit 60 supplies a necessary solution to the pores 14 for each inspection process.
  • the cavity 16 is not open on the other surface 12 side, so that the supplied solution does not enter the cavity 16 from the other surface 12 side.
  • the solution supply unit 60 supplies the solution so that the solution does not enter the cavity from one surface 11 where the cavity 16 is opened. Control. As shown in FIG.
  • the solution supply unit 60 was provided with a lid portion 67 that covers the opening of the cavity portion 16 in a part of the depressurizing space portion 66, and overflowed from the opening of the pores 14 on one surface 11.
  • the configuration may be such that the solution does not enter the cavity 16.
  • the light emitted from the inspection device and detected by the photodetector is, for example, fluorescence generated when the label attached to the test substance is excited, or luminescence generated by the label acting as a substrate of the reaction solution It is an optical signal resulting from a label, such as luminescence by reaction (hereinafter referred to as “chemiluminescence”).
  • chemiluminescence an optical signal resulting from a label, such as luminescence by reaction
  • the optical signal includes absorbance (colorimetric) in addition to fluorescence and chemiluminescence light.
  • FIG. 14 is a diagram schematically showing the inspection process.
  • a capturing substance 30 such as an allergen is fixed to the inner wall surface 14a of the pore 14 of the inspection device 1 (S1).
  • a specimen liquid containing a test substance (for example, a specific IgE antibody that specifically binds to the allergen) is supplied to the pores 14 of the test device 1, and the test substance 32 is bound to the capture substance 30 (S2). ).
  • the above-described solution supply unit 60 is used for supplying the sample liquid.
  • the sample liquid is efficiently brought into contact with the capture substance 30 in the pores 14 by repeatedly pumping and discharging the sample liquid by the pipette unit 64 while the sample liquid is stored in the storage unit 62. Can do.
  • a labeled solution containing a labeled substance 35 in which a label F is added to a substance 33 (for example, a secondary antibody) that specifically binds to the test substance 32 is supplied to the pores 14.
  • a substance 33 for example, a secondary antibody
  • the labeling substance 35 is bound to the test substance 32 (S3).
  • the label solution is supplied to the pores 14 by the solution supply unit 60 in the same manner as the sample solution.
  • the labeling substance 35 is bound to the test substance 32 after the test substance 32 is bound to the capture substance 30.
  • the sample liquid and the labeling solution are mixed in advance outside the test apparatus 50.
  • the test substance 32 and the labeling substance 35 may be combined to supply the mixed liquid into the pores 14.
  • the test substance 32 to which the labeling substance 35 is bound can be bound to the capture substance 30 fixed in the pores 14.
  • the cleaning solution is supplied to the pores 14 using the solution supply unit 60 in the same manner as the supply of the sample solution, and nonspecific adsorption is performed in the pores 14.
  • the test substance 32 and the labeling substance 35 are removed.
  • the cleaning liquid is discharged, and the optical signal from the luminescence reaction in which the label F acts as a catalyst is detected by the photodetector 70 in a state where the inspection solution such as the buffer solution is filled in the pores 14 ( S4).
  • the inspection solution such as the buffer solution is filled in the pores 14 ( S4).
  • the buffer solution is supplied into the pore 14 as a test solution, and the fluorescence measurement is performed with the pore 14 filled with the buffer solution. .
  • the inspection device is irradiated with light having a wavelength that excites the fluorescent label as excitation light, and fluorescence from the label excited by the excitation light is detected.
  • the photodetector 70 used for detecting the fluorescent label is provided with an excitation light irradiation unit.
  • the label F is an enzyme or the like that acts as a catalyst for a chemical reaction in which luminol is oxidized by hydrogen peroxide to generate luminescence
  • the chemical by contacting with the enzyme that acts as a catalyst after the washing treatment.
  • a reaction solution containing a substance that emits light by the reaction is supplied into the pores 14 as a test solution.
  • the photodetector 70 by supplying this reaction solution to the pores 14, the enzyme label imparted to the test substance 32 captured by the capture material 30 in the pores 14 becomes the substance in the reaction solution.
  • Luminescence generated by catalyzing a chemical reaction is detected. In this case, luminescence detection is performed in a state where the pores 14 are filled with the reaction solution.
  • enzyme labels that generate chemiluminescence include enzymes that react with chemiluminescent substrates such as luminol, lophine, lucigenin and oxalate.
  • HRP horseradish peroxidase
  • a reaction liquid luminol reaction liquid
  • HRP functions as a catalyst an enzyme for converting HRP to alkaline phosphatase. It is preferable to use a reaction solution containing a dioxetane chemiluminescent substrate.
  • the luminol reaction solution contains at least a luminol substrate and a hydrogen peroxide solution.
  • the enzyme label catalyzes the oxidation of luminol in the presence of hydrogen peroxide.
  • the reaction solution preferably contains a sensitizer that sensitizes chemiluminescence.
  • chemiluminescent substrate described above but also a reaction solution containing a luminescent substrate or a fluorescent substrate may be used to detect a color (absorption) reaction or fluorescence. it can.
  • the inspection device of the present invention by using the inspection device of the present invention, the light generated in the pores is efficiently emitted from the surface of the inspection device, and the light extraction efficiency is high. Is possible.

Abstract

Le problème décrit par la présente invention est de fournir un dispositif d'inspection qui peut améliorer l'efficacité d'extraction de lumière et qui peut être fabriqué à un faible coût, en plus d'un appareil d'inspection équipé du dispositif d'inspection, et un procédé d'inspection. La solution selon l'invention concerne un dispositif d'inspection (1) comprenant une pluralité de pores (14) qui traversent un substrat en forme de plaque (10) d'une première surface (11) à l'autre surface (12), et une cavité (16) qui s'étend dans la direction de profondeur des pores (14), qui ne pénètre pas au moins dans la seconde surface et est formée de façon à entourer une zone (20) qui comprend au moins certains de la pluralité de pores (14) lorsqu'ils sont vus depuis la première surface (11), une partie du substrat en forme de plaque (10) qui constitue une zone (20) comprenant au moins certains des pores (16) transmettant la lumière, et une substance fixatrice (30) qui se lie spécifiquement à une substance spécifique étant immobilisée sur la surface de paroi interne (14a) d'au moins certains des pores (14).
PCT/JP2017/010091 2016-03-30 2017-03-14 Dispositif d'inspection, appareil d'inspection et procédé d'inspection WO2017169714A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
JP2016-067479 2016-03-30
JP2016067479 2016-03-30

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WO2017169714A1 true WO2017169714A1 (fr) 2017-10-05

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Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2003066042A (ja) * 2001-08-27 2003-03-05 Nippon Shokubai Co Ltd バイオチップ
JP2004286579A (ja) * 2003-03-20 2004-10-14 Kanagawa Acad Of Sci & Technol Ｄｎａ分析用アレイおよびそれを用いたｄｎａ分析システムおよび分析方法
JP2005283556A (ja) * 2004-03-05 2005-10-13 Canon Inc 標的物質認識素子、検出方法及び装置
JP2006038754A (ja) * 2004-07-29 2006-02-09 Fuji Photo Film Co Ltd 標的検出用基材及びその製造方法、並びに、標的検出装置及び標的検出方法
JP2006105879A (ja) * 2004-10-07 2006-04-20 Japan Science & Technology Agency 分子認識チップとその製造方法、分子認識チップ分析方法並びに顕微赤外ｄｎａ解析装置

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2003066042A (ja) * 2001-08-27 2003-03-05 Nippon Shokubai Co Ltd バイオチップ
JP2004286579A (ja) * 2003-03-20 2004-10-14 Kanagawa Acad Of Sci & Technol Ｄｎａ分析用アレイおよびそれを用いたｄｎａ分析システムおよび分析方法
JP2005283556A (ja) * 2004-03-05 2005-10-13 Canon Inc 標的物質認識素子、検出方法及び装置
JP2006038754A (ja) * 2004-07-29 2006-02-09 Fuji Photo Film Co Ltd 標的検出用基材及びその製造方法、並びに、標的検出装置及び標的検出方法
JP2006105879A (ja) * 2004-10-07 2006-04-20 Japan Science & Technology Agency 分子認識チップとその製造方法、分子認識チップ分析方法並びに顕微赤外ｄｎａ解析装置

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