WO2017169717A1 - 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
WO2017169717A1
WO2017169717A1 PCT/JP2017/010100 JP2017010100W WO2017169717A1 WO 2017169717 A1 WO2017169717 A1 WO 2017169717A1 JP 2017010100 W JP2017010100 W JP 2017010100W WO 2017169717 A1 WO2017169717 A1 WO 2017169717A1
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Prior art keywords
inspection device
substance
inspection
pores
solution
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PCT/JP2017/010100
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English (en)
Japanese (ja)
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弘隆 渡野
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富士フイルム株式会社
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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
    • G01N33/551—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals the carrier being inorganic
    • G01N33/553—Metal or metal coated

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 discloses a porous substrate having a large number of through holes in an aluminum (Al) base material. Since aluminum has a higher reflectance than silicon, it is considered that the light extraction efficiency can be improved as compared with the case where a silicon substrate is used. However, it cannot be said that the light extraction efficiency can be sufficiently improved because the light exit openings of the individual pores are small. On the other hand, if the pore diameter is increased in order to widen the light extraction opening, the pore density is lowered, and the amount of trapped substance that can be fixed as a whole is reduced.
  • Patent Document 2 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.
  • JP 2004-93152 A Japanese Patent No. 4125244
  • Patent Document 2 Although the light extraction efficiency can be improved as the optical aperture ratio increases, the reflectance of silicon used as a frame wall is low, so that light incident on the silicon wall is also attenuated. Therefore, the extraction efficiency of light incident on the frame wall is not improved.
  • it is considered necessary to increase the thickness of the chip, expand the detection area, or increase the density of the pores.
  • 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.
  • 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 region including at least a part of the pores, wherein at least a part of the wall member constituting the inner wall surface of the at least a part of the pores is made of alumina, and when viewed from the one side, A frame wall made of aluminum surrounding the region,
  • a capture substance that specifically binds to a specific substance is fixed to an inner wall surface of the at least some pores.
  • the wall member has a fine hole extending in parallel with the pore and having a diameter smaller than the pore.
  • the wall member is preferably made of anodized alumina.
  • the frame wall is preferably provided in contact with the wall member.
  • the other surface is preferably made of aluminum.
  • the capture substance is preferably an antigen, an antibody, or deoxyribonucleic acid (DNA).
  • the inspection device of the present invention comprises a plurality of the above-mentioned regions surrounded by a frame wall made of aluminum, and the same type of capture material is bound as a capture material to the inner wall surface of the pore in one of the plurality of regions. It is preferable that the plurality of regions have regions in which the bound capture substances are different from each other.
  • 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 fluorescence 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 label
  • the light emitted from the inspection device is as follows: You may detect the light which a substrate in a reaction liquid produces when catalyzed by an enzyme label
  • 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.
  • 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, and is an area including at least some of the pores, Since at least a part of the wall member constituting the inner wall surface of some of the pores has a region made of alumina and a frame wall made of aluminum surrounding the region when viewed from the one surface, The extraction efficiency of the light generated in the pores to at least one surface side of the inspection device can be improved.
  • 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 is a wall made of alumina that constitutes an inner wall surface 14a of a pore in a plate-like substrate 10 having a plurality of pores 14 penetrating from one surface 11 to the other surface 12. It has the area
  • FIG. In the present embodiment a total of nine pores 14 in the vertical and horizontal directions are used as one region 20, and a frame wall 18 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. 4 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.
  • the wall member constituting the inner wall surface 14a of the four regions 20 is made of alumina except for a part on the other surface 12 side.
  • a frame wall 18 made of aluminum is provided surrounding the region 20 and in contact with the wall member 16 made of alumina.
  • the lower surface member 19 which comprises the other surface 12 of the plate-shaped base material 10 consists of aluminum.
  • the lower surface member 19 is continuous with the frame wall 18 and is integrally formed.
  • the plate-like substrate 10 is composed of aluminum and alumina over the entire region.
  • the lower surface member 19 made of aluminum, a component of the light generated in the pores 14 that is emitted to the other surface 12 side and incident on the lower surface member 19 is reflected to the one surface 11 side. Therefore, the light extraction efficiency to the one surface 11 side can be improved.
  • the inspection device may not have the lower surface member 19.
  • the same result can be obtained even if the light detection is performed from either one of the surfaces 11 side and the other surface 12 side.
  • the wall member constituting the inner wall surface 14a of the region 20 including at least a part of the pores 14 is made of alumina. That is, in the region 20 surrounded by the frame wall 18, the wall member may include a portion formed of alumina and a portion formed of another material (for example, aluminum).
  • the frame wall 18 is formed so as to surround the outer periphery of the region 20, but is partially cut off if the length of the unconnected portion is about 20% of the length of the outer periphery of the region 20. It may be.
  • the plate-like base material 10 is preferably one in which at least a part of an aluminum base material is oxidized to form a wall member 16 made of alumina. According to such a configuration, an inexpensive aluminum base material can be used, and since it can be easily manufactured, it is preferable because it can be manufactured at low cost.
  • the plate-like substrate 10 may include a portion made of a different material other than aluminum and alumina.
  • 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.
  • the region 20 surrounded by the frame wall 18 (hereinafter referred to as “inspection region 20”) may include only one pore 14, but a plurality of pores 14 are included. It is preferable to include.
  • 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).
  • the capture substance 30 that is fixed to the inner wall surface 14a of the pore 14 and specifically binds to a specific substance is a substance that specifically binds to these test substances.
  • the test device 1 is suitable for an allergy test provided with an allergen that is a kind of antigen as a capture substance.
  • the present inspection device 1 has a plurality of inspection regions 20 partitioned by a frame wall 18.
  • a trapping substance of the same type is bound (fixed) to the inner wall of the pore 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.
  • the plurality of inspection regions there may be two or more regions to which the same type of capture substance is bound.
  • inspection regions 20 in which different capture substances are fixed may be alternately arranged, and inspection regions in which the same type of capture material is periodically fixed may be disposed. According to such a configuration, it is possible to suppress variation and obtain a highly reliable test result.
  • a plurality of allergen test regions 20 are provided by fixing the same type of allergen as a capture substance in one or a plurality of test regions 20 in one test device. It is preferable that the response to can be simultaneously examined.
  • FIG. 6 is an enlarged perspective view showing a part of the inspection device 1.
  • the wall member 16 made of alumina preferably has fine holes 17 extending in parallel with the pores 14 and having a diameter smaller than the pores 14.
  • Such fine holes 17 can be formed by anodizing aluminum to transform it into alumina.
  • porous alumina having a large number of aligned fine holes 17 can be easily obtained.
  • the diameter of the fine hole formed by anodic oxidation is about several nm to several tens of nm, and preferably 5 nm to 20 nm.
  • the fine hole 17 is preferably not a through-hole penetrating from one surface 11 to the other surface 12 but a non-through hole with at least the other surface 12 closed.
  • a non-through hole can be formed by providing the lower surface member 19 described above.
  • a barrier layer (dense alumina layer) formed between the base material base and the anodizing treatment may be provided.
  • the one surface 11 side may remain open, it is also preferable that a sealing process for closing the opening of the one surface 11 is performed.
  • the fine hole 17 is made into the closed cavity part 17a.
  • the hollow portion 17a formed in this way can also be regarded as a kind of fine hole. According to such a configuration, the infiltration of the solution from any one of the one surface 11 and the other surface 12 can be suppressed, and the cavity can be used as an air layer, contributing to the improvement of light extraction efficiency described later. Becomes easy.
  • FIG. 7 is a schematic view showing a part of the inspection device 1 in an enlarged manner.
  • the present inspection device 1 binds a test substance to a capture substance (not shown in FIG. 7) fixed to the inner wall surface 14a of the pore 14, and specifically detects the test substance.
  • a binding substance labeled with a binding enzyme By binding a binding substance labeled with a binding enzyme and detecting a light signal generated by a luminescence reaction in which this enzyme acts as a catalyst, the presence or absence of the test substance or the amount of the test substance is detected. It is used in an inspection method for detecting the presence or absence of a test substance and the amount of a 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.
  • the wall member 16 constituting the inner wall surface 14a of the pore 14 in the inspection region 20 is made of alumina and has light transmittance.
  • a part of light generated in one pore 14 is repeatedly reflected in one pore and emitted from the surface.
  • the wall member 16 has optical transparency, a part of the light generated in one pore 14 can be guided to the surface side through the adjacent pore. As a result, the aperture ratio of light emitted to the surface of the light generated in the pores 14 increases.
  • the frame wall 18 made of aluminum is provided so as to surround the inspection region 20, a part of the light generated in the pores 14 in the inspection region 20 is reflected by the interface C 0 with the frame wall 18 and is a plate. The light is guided to the surface side of the substrate. Since aluminum has a higher reflectance than silicon and is less attenuated by reflection, more light components can be extracted to the device surface.
  • the lower surface member 19 constituting the other surface 12 is made of aluminum as in the inspection device 1 of the present embodiment, the light that is generated in the pore 14 and directly or indirectly incident on the lower surface member 19. Can be reflected on the one surface 11 side. In this case, the light emitted to the one surface 11 side can be further increased, and the extraction efficiency can be further improved.
  • FIG. 8 is an enlarged view of a region VIII indicated by a broken line in FIG.
  • the fine holes 17 provided in the wall member 16 are used so that the solution 61 does not enter, and the fine holes 17 constitute an air layer.
  • the wall member 16 has a multi-layered structure of the wall member layer 16a and the air layer (the minute hole 17) microscopically as shown in FIG.
  • the wall member layer 16a has a refractive index of about 1.6 to 1.9, and the inspection solution 61 has a refractive index 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 wall member 16 and the air layer is very large, at least a part of the light incident on the interface between the wall member layer 16a and the air layer is reflected at the interface. Since the wall member layer 16a has a larger refractive index than the air layer, the light incident on the interface C 1 at an incident angle theta 1 larger than the critical angle from the wall member 16 side to the air layer is totally reflected, a plate-shaped base It will go to the surface side of the material 10.
  • the light generated from the pores 14 in the region 20 can be reflected by the frame wall 18 made of aluminum having high reflectivity and guided to the surface side. Light attenuation can be suppressed and light extraction efficiency can be improved. Furthermore, when the micro hole 17 is provided in the wall member 16, light is condensed on the surface side of the device by causing reflection at a plurality of interfaces by the wall member 16, the air layer, and the multi-layered structure. Therefore, the light extraction efficiency can be greatly improved as compared with the case where a frame made of silicon disclosed in Patent Document 2 is provided.
  • FIG. 9 is a diagram illustrating a manufacturing process of the inspection device 1.
  • an aluminum substrate 100 is prepared (A1).
  • a mask 102 is formed on a portion to be the frame wall 18 made of aluminum on one surface and the entire surface of the other surface (A2).
  • the mask can be formed by a known lithography method or screen printing. For example, a method described in JP 2014-198353 A can be employed.
  • the aluminum base material 100 provided with the mask 102 at the part where the frame wall is to be formed is anodized.
  • known anodic oxidation methods described in JP-A-2015-132837 and JP-A-2014-198453 can be employed.
  • the portion of the aluminum substrate 100 that is not provided with the mask 102 is anodized to form an anodized alumina portion 104 (A3).
  • a mask is removed (A4).
  • the pores 14 are formed in the anodized alumina portion 104 (A5).
  • the pores 14 can be formed by a mechanical method using a drill or the like.
  • a mask may be provided in a portion other than the portion where the pore 14 is formed in the anodized alumina portion 104, and the pore 14 may be formed by chemical etching.
  • the method for producing the plate-like substrate 10 is not limited to the above procedure, and anodization is performed by providing the aluminum substrate with pores 14 first, and then providing a mask at the frame wall forming portion. It may be a procedure.
  • the anodized alumina portion 104 formed in the above procedure A3 is porous alumina and has a large number of fine holes. By performing a wide process on the desired fine holes, the diameter is larger than that of the fine holes. Large pores 14 can also be formed.
  • a known method can be used as the power wide processing, and for example, a method described in Japanese Patent Application Laid-Open No. 2014-198453 can be employed.
  • the other surface 12 is composed of a lower surface member 19 made of aluminum, and the fine hole 17 does not penetrate to the other surface 12 side.
  • An enlarged view of a region X indicated by a broken line of the plate-like substrate 10 shown in A5 of FIG. 9 is shown in A5 of FIG.
  • the barrier layer 101 and the aluminum substrate 100 that is the substrate substrate remain between the micro holes 17 and the other surface 12.
  • the other surface 12 side can be a closed hole.
  • the lower surface member 19 is comprised by the base-material base
  • the fine hole 17 can be formed as a non-through hole closed on the other surface 12 side without penetrating on the other surface 12 side.
  • the sealing treatment include high-temperature pressurized steam treatment and boiling treatment in boiling water.
  • the alumina on the surface becomes a hydrate, and the opening is substantially closed by the alumina hydrate layer 16b.
  • FIG. 10A5 the opening is shown as being completely blocked, but it is sufficient that the opening is blocked to the extent that the solution can be prevented from entering, and the opening is formed by the alumina hydrate layer 16b. It does not have to be completely blocked.
  • the capture substance which is a specific binding substance that specifically binds to the test substance is bound (immobilized) to the inner wall surface 14a of the pore 14 of the plate-like substrate 10 manufactured as described above.
  • the inspection device 1 can be manufactured.
  • a known method can be applied without any particular limitation.
  • the method disclosed in the above-described Patent Document 2 can be used.
  • FIG. 11 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.
  • FIG. 12 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 fine holes 17 (not shown in FIG. 12) provided in the wall member 16 are non-open on the other surface 12 side, so that the supplied solution is fine holes from the other surface 12 side. 17 does not enter.
  • the solution supply unit 60 supplies the solution so that the solution does not enter the fine hole 17 from the one surface 11 where the fine hole 17 is opened. Control. If the above-described sealing treatment is applied to the opening on the one surface 11 side of the fine hole 17, the solution does not enter the fine hole 17 from the one surface 11. Is preferable.
  • the light emitted from the test device and detected by the photodetector is, for example, fluorescence generated by excitation of a label attached to the test substance, or binding of an antibody that specifically binds to the test substance.
  • the substance is given the same label as above, and light is emitted from the label, or an enzyme is labeled on a binding substance such as an antibody that specifically binds to the test substance, and light emission by the reaction using this enzyme as a catalyst (in the following) , Or “chemiluminescence”)).
  • chemiluminescence if the test substance generates autofluorescence, no label is necessary, and autofluorescence may be detected.
  • the optical signal includes absorbance (colorimetric) in addition to fluorescence and chemiluminescence light.
  • FIG. 13 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. It is also possible to combine the test substance 32 and the labeling substance 35 by mixing and supply the mixed liquid into the pores 14. In this case, by supplying the mixed liquid to 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 that reacts with the label F as a catalyst is detected by the photodetector 70 with the inspection solution such as the buffer solution filled in the pores 14 ( S4).
  • the wall member 16 is transparent and has a frame wall 18 around it. Since the air layer 17 constitutes an air layer, light detection can be performed efficiently.
  • 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 the reaction solution is supplied to the pores 14 so that the chemiluminescent label attached to the test substance 32 captured by the capture material 30 in the pores 14 is a substance in the reaction solution.
  • Luminescence generated by reaction with 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 permettant d'améliorer l'efficacité d'extraction de la lumière et pouvant être produit à faible coût ; un appareil d'inspection équipé dudit dispositif d'inspection ; et un procédé d'inspection. La solution de l'invention porte sur un dispositif d'inspection (1) pourvu d'une pluralité de trous (14) qui s'étendent d'une surface (11) à l'autre surface (12) d'un substrat de type feuille (10), ledit dispositif d'inspection (1) comprenant : des zones (20), chacune d'entre elles contenant au moins certains des trous (14) et au moins une partie des éléments de paroi (16) qui constituent les surfaces de paroi intérieure (14a) desdits au moins certains trous (14) comprenant de l'alumine; et des parois d'encadrement (18) comprenant de l'aluminium, lesquelles entourent lesdites zones (20) lorsque le dispositif est vu depuis la surface (11). Une substance de piégeage qui se lie spécifiquement à une substance spécifique est fixée sur les surfaces de paroi intérieure (14a) desdits au moins certains trous (14).
PCT/JP2017/010100 2016-03-30 2017-03-14 Dispositif d'inspection, appareil d'inspection et procédé d'inspection WO2017169717A1 (fr)

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JP2016-067482 2016-03-30

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Citations (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2000229228A (ja) * 1999-02-10 2000-08-22 Nippon Telegr & Teleph Corp <Ntt> 多孔質メンブレンおよびその作製方法
JP2003066042A (ja) * 2001-08-27 2003-03-05 Nippon Shokubai Co Ltd バイオチップ
JP2004093152A (ja) * 2002-08-29 2004-03-25 Fuji Photo Film Co Ltd 生化学解析用ユニットを利用した化学発光法
JP2005106671A (ja) * 2003-09-30 2005-04-21 Fuji Photo Film Co Ltd 生化学解析用ユニットを用いた反応方法
JP2007163163A (ja) * 2005-12-09 2007-06-28 Sharp Corp バイオケミカルセンサ及び測定装置
JP2008076313A (ja) * 2006-09-22 2008-04-03 Japan Advanced Institute Of Science & Technology Hokuriku 分析用チップ及びその製造方法、分析装置並びに分析方法
JP2010019570A (ja) * 2008-07-08 2010-01-28 Funai Electric Advanced Applied Technology Research Institute Inc マトリックスセンサ及びセンサシステム
WO2013179342A1 (fr) * 2012-05-30 2013-12-05 株式会社バイオマーカーサイエンス Biopuce

Patent Citations (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2000229228A (ja) * 1999-02-10 2000-08-22 Nippon Telegr & Teleph Corp <Ntt> 多孔質メンブレンおよびその作製方法
JP2003066042A (ja) * 2001-08-27 2003-03-05 Nippon Shokubai Co Ltd バイオチップ
JP2004093152A (ja) * 2002-08-29 2004-03-25 Fuji Photo Film Co Ltd 生化学解析用ユニットを利用した化学発光法
JP2005106671A (ja) * 2003-09-30 2005-04-21 Fuji Photo Film Co Ltd 生化学解析用ユニットを用いた反応方法
JP2007163163A (ja) * 2005-12-09 2007-06-28 Sharp Corp バイオケミカルセンサ及び測定装置
JP2008076313A (ja) * 2006-09-22 2008-04-03 Japan Advanced Institute Of Science & Technology Hokuriku 分析用チップ及びその製造方法、分析装置並びに分析方法
JP2010019570A (ja) * 2008-07-08 2010-01-28 Funai Electric Advanced Applied Technology Research Institute Inc マトリックスセンサ及びセンサシステム
WO2013179342A1 (fr) * 2012-05-30 2013-12-05 株式会社バイオマーカーサイエンス Biopuce

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