WO2017169715A1 - 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
WO2017169715A1
WO2017169715A1 PCT/JP2017/010093 JP2017010093W WO2017169715A1 WO 2017169715 A1 WO2017169715 A1 WO 2017169715A1 JP 2017010093 W JP2017010093 W JP 2017010093W WO 2017169715 A1 WO2017169715 A1 WO 2017169715A1
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Prior art keywords
inspection device
inspection
pores
substance
hollow portion
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PCT/JP2017/010093
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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 WO2017169715A1 publication Critical patent/WO2017169715A1/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 a label that emits fluorescence upon receiving excitation light, or a label that catalyzes a substrate reaction to generate color, fluorescence, or chemiluminescence. Or the like 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 amount of extracted light is increased by making the diameter of the pores of the porous substrate larger than the inside in the diameter of the opening portions on the front and back sides.
  • Patent Document 3 proposes a device having tapered pores whose diameter gradually increases toward the light extraction surface of the substrate.
  • Patent Document 1 Although it is possible to improve the light extraction efficiency with an increase in the light aperture ratio, the light incident on the silicon wall is attenuated because the reflectance of silicon used as the frame wall is low. The extraction efficiency of light incident on the frame wall is not improved.
  • Patent Document 2 since the opening diameter on both surfaces of one side and the other surface of porous silicon is larger than the inner opening diameter, the light traveling toward one surface or the other surface side The light extraction efficiency is increased because the light aperture ratio is improved. However, since light that travels to one surface or the other surface is not directed to the opposite surface, a light detector is disposed only on one surface or only on the other surface. When used in a measuring apparatus that performs detection, it is not possible to achieve a sufficient improvement in extraction efficiency.
  • Patent Document 3 a portion of light that has occurred in the pores and that has progressed toward the surface opposite to the light extraction surface is formed by tapered pores whose diameter gradually increases toward the light extraction surface of the substrate. Although it can be reflected by the inner wall surface and directed toward the light extraction surface side, it is expressed by the ratio of the length of the pore to the diameter of the pore due to the demand for higher performance of the chip as described above. When the aspect ratio becomes large, a sufficient taper angle cannot be set, so that the light extraction efficiency cannot be sufficiently improved.
  • 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,
  • the pore is continuous with the columnar hollow portion extending with the same opening diameter and the inner wall surface forming the columnar hollow portion formed on the other surface side of the columnar hollow portion, and the pore is larger than the opening diameter in the columnar hollow portion.
  • Having a reflective surface to reduce the opening diameter of This is an inspection device in which the angle formed by the reflection surface and the other surface is 0 ° or more and 45 ° or less.
  • a trapping substance that specifically binds to a specific substance is fixed on the inner wall surface of the hollow columnar portion of the present invention.
  • the capture substance is preferably an antigen, an antibody, or deoxyribonucleic acid (DNA).
  • the reflective surface is preferably a metal surface.
  • the pore has a tapered portion that gradually decreases the opening diameter of the pore from one surface toward the columnar hollow portion.
  • the plate-like substrate may be made of one or more materials of Si, SiO 2 , Al, Al 2 O 3 and a resin material.
  • the material constituting the reflecting surface may be different from the material constituting the columnar hollow portion.
  • the opening diameter in the columnar hollow portion of the pore is preferably 1 ⁇ m to 100 ⁇ m.
  • the opening diameter is defined by the equivalent circle diameter of the opening region in a cross section parallel to one surface.
  • the equivalent circle diameter refers to the diameter of a circle having the same area as the area of the opening region.
  • 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 side of the inspection device and detects light emitted from the inspection device.
  • the inspection method of the present invention fixes a capture substance that specifically binds to a specific substance on the inner wall surface of the columnar hollow portion of the at least some pores of the inspection device of the present invention, Supply the sample liquid containing the specific substance, and bind the specific substance to the capture substance.
  • a labeling substance that specifically binds to a specific substance is bound to the specific substance, This is an inspection method for detecting light emitted from the one surface of the inspection device in a state in which the inspection solution is supplied to the pores and the inspection solution is retained in the pores.
  • an enzyme label is used as the label
  • the light produced when the substrate 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, excitation light that excites the fluorescent label is irradiated to the inspection device, and the labeling substance is irradiated by excitation light as light emitted from the inspection device. Fluorescence resulting from may be detected.
  • the 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, and the columnar hollow portion in which the pores extend with the same opening diameter, and the columnar shape
  • the reflective surface formed on the other surface side of the hollow portion is continuous with the inner wall surface of the columnar hollow portion and has a smaller opening diameter than the pores of the columnar hollow portion. Since the formed angle is 0 ° or more and 45 ° or less, it is possible to improve the extraction efficiency of light generated in the pores to the one surface side of the inspection device.
  • FIG. 1 It is a perspective view of the inspection device concerning the 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 includes a plate-like base material 10 having a plurality of pores 14 penetrating from one surface 11 to the other surface 12, and the pores 14 extend in the same opening diameter.
  • 40 and a reflecting surface that is formed on the other surface 12 side of the columnar hollow portion 40 and that is continuous with the inner wall surface 41 constituting the columnar hollow portion 40 and that makes the opening diameter ⁇ of the pore 14 smaller than that of the columnar hollow portion 40. 42.
  • the reflection surface 42 has an angle ⁇ with the other surface 12 of 0 ° or more and 45 ° or less.
  • the inner wall surface 14 a of the pore 14 includes at least an inner wall surface 41 and a reflecting surface 42 that constitute the columnar hollow portion 40.
  • a trapping substance 30 that specifically binds to a specific substance is fixed to the inner wall surface 14 a of the pore 14.
  • acquisition substance 30 should just be provided in the inner wall surface 41 which comprises a columnar hollow part among the inner wall surfaces 14a.
  • 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 as shown in FIG. 1, in the following, one surface 11 and the other surface 12 of the plate-like base material 10 are Sometimes referred to as one surface and the other surface of the inspection device 1.
  • the plate-like substrate 10 is made of one or more materials of Si (silicon), SiO 2 (silicon oxide), Al (aluminum), Al 2 O 3 (alumina), and a resin material. Preferably it is.
  • the plate-like substrate 10 may be opaque made of Si or Al, or may be transparent made of SiO 2 or Al 2 O 3 .
  • the plate-like substrate 10 may be composed of any single material, or may include, for example, a Si portion and a SiO 2 portion by partially oxidizing the Si substrate,
  • the Al base material may be partially oxidized to include an Al portion and an Al 2 O 3 portion.
  • the plate-like substrate 10 may be composed of a main body member made of Si, SiO 2 , Al, or Al 2 O 3 and a surface member made of a resin material.
  • the thickness t (see FIG. 3) of the plate-like substrate 10 is not particularly limited, but is preferably about 100 ⁇ m to 2000 ⁇ m.
  • the length (thickness) t 1 of the columnar hollow portion 40 and the length t 2 in the pore length direction of the portion where the reflecting surface 42 is arranged in a tapered shape hereinafter, “ The total of the thickness t 2 of the reflecting surface 42 is called the thickness t 2 of the plate-like substrate 10.
  • the pores 14 provided in the plate-like substrate 10 are preferably arranged as shown in the present embodiment, but may be arranged randomly. In the present embodiment, 9 pores of 3 rows ⁇ 3 columns are provided for easy visual recognition, but the inspection device 1 includes, for example, 100 pores of 10 rows ⁇ 10 columns. There is no limit to the number of pores provided in the case. In addition, for example, an inspection device including a plurality of inspection regions may be provided with 100 pore regions of 10 rows ⁇ 10 columns as one inspection region.
  • the cross-sectional shape (opening shape) parallel to the one surface 11 of the pore 14 is a square shape, but the opening shape is not limited to a rectangle such as a square or a rectangle, but a circle, an ellipse, a triangle, It may be a pentagon or more polygon. Note that the corners of the polygon may be rounded due to manufacturing reasons.
  • the opening diameter ⁇ of the pore 14 is a circle-equivalent diameter in a cross-sectional shape (opening region) parallel to one surface 11 of the pore 14.
  • the opening diameter ⁇ of the pore 14 is the same in the columnar hollow portion 40, and is configured to become smaller as it approaches the other surface 12 by the reflecting surface 42.
  • the opening diameter in the columnar hollow portion 40 of the pores 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 aspect ratio (t / ⁇ ) represented by the thickness of the plate-like substrate with respect to the opening diameter in the columnar hollow portion 40 is preferably 5 or more and 100 or less. When the aspect ratio is within this range, the effect of providing the reflecting surface 42 is particularly remarkable.
  • the aspect ratio with respect to the opening diameter of the thickness from the one surface 11 side with respect to the opening diameter in the columnar hollow part 40 to the portion including the reflecting surface 42 is 5 or more and 100 or less. It is preferable that
  • the reflection surface 42 only needs to be reflective to light generated due to a sign described later.
  • having the reflectivity means that the reflectance with respect to the light to be reflected is 50% or more.
  • the reflectance at least for the wavelength of 550 nm may be 50% or more.
  • a reflectance of 50% or more with respect to the entire visible light (wavelength 400 nm to 700 nm) is preferable because it can be applied regardless of the type of label.
  • the reflective surface 42 may be a metal surface made of a metal such as steel, stainless steel, aluminum, gold, platinum, silver, copper, titanium, nickel, cobalt, tin, tungsten, molybdenum, or niobium.
  • high resin reflectance such as white resin, for example, may be composed of pigments or fillers such as TiO 2, Al 2 O 3.
  • the angle ⁇ between the reflecting surface 42 and the other surface 12 is 0 ° to 45 °.
  • ⁇ is 0 °
  • the reflecting surface 42 and the other surface 12 are parallel.
  • the reflecting surface 42 is provided as a tapered portion that gradually decreases the opening diameter of the pore from the columnar hollow portion 40 toward the other surface 12.
  • FIG. 5 is a schematic view showing a part of the cross section of the inspection device 1.
  • This test device 1 binds a test substance to a capture substance (not shown in FIG. 5) fixed to the inner wall surface 14a of the pore 14, and further receives a secondary antibody labeled with an enzyme. Used in a test method for detecting the presence or absence of the test substance and / or the amount of the test substance by detecting an optical signal that is chemiluminescent using the enzyme (label F) as a catalyst by binding to the test substance. It is done.
  • the light generated in one pore 14 is repeatedly reflected by the inner wall surface 14a of the pore 14 and emitted from the surface.
  • the reflecting surface 42 by providing the reflecting surface 42, the light generated by the sign F and radiated to the other surface 12 side is reflected directly or by the inner wall surface 14a and incident on the reflecting surface 42 is reflected.
  • the light is reflected by the surface 42 and guided to the one surface 11 side. As a result, the amount of light emitted from the one surface 11 of the light generated in the pores 14 can be effectively increased.
  • the reflecting surface 42 having an angle ⁇ formed with the other surface 12 of 45 ° or less is continuously provided on the inner wall surface 41 constituting the columnar hollow portion 40, the inner wall surface 14 a of the pore 14, in particular, Of the light generated on the inner wall surface 41 of the columnar hollow portion 40, the light incident on the reflecting surface 42 can be efficiently reflected. If the angle ⁇ is 45 ° or less, the proportion of the light incident on the reflective surface 42 that is reflected toward one surface is set to be equal to or greater than the proportion of the light reflected toward the other surface. Therefore, the effect of improving the light extraction amount on the one surface 11 side is high. As the angle ⁇ approaches 0 °, it is possible to increase the proportion of light reflected toward one surface of the light incident on the reflecting surface 42. However, when the angle ⁇ is small, the opening diameter on the other surface 12 is relatively small, and it may be difficult to suck the test solution from the other surface 12 into the pores. What is necessary is just to determine suitably in the range of -45 degrees.
  • the light extracted from the one surface 11 side is significantly increased by reflecting the light generated from the pores 14 by the reflection surface 42 provided on the other surface 12 side. Can be made.
  • 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.
  • different trapping substances can be fixed between the plurality of pores 14. 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 pores containing the same type of trapping substance between the plurality of pores 14. If an inspection device in which pores 14 in which different capture substances are fixed are alternately arranged and pores in which the same kind of capture substance is fixed is used is used, a test result with reduced variation is obtained. It becomes possible.
  • a plurality of allergens can be provided in one test device by providing pores to which different types of allergens are fixed. It becomes possible to test the response to
  • a capture substance may be bound (fixed), and different capture substances may be fixed between a plurality of examination regions. Since a plurality of pores are provided in one inspection region, it is possible to simultaneously inspect a plurality of test substances while improving the sensitivity to one kind of test substance. Note that there may be two or more regions containing the same type of capture substance between a plurality of inspection regions.
  • FIGS. 6 to 10 are sectional views of a part of the inspection device of the design change example.
  • a plurality of pores 14 shown in each figure are arranged two-dimensionally.
  • the reflection surface 43 is parallel to the other surface 12 of the plate-like substrate 10, that is, the angle formed with the other surface is 0 °.
  • the pore 14 has an inner wall surface 14 a composed of an inner wall surface 41 of the columnar hollow portion 40, a reflecting surface 43, and an inner wall surface 48 constituting a small-diameter hollow portion having an opening diameter smaller than that of the columnar hollow portion 40.
  • the total of the thickness t 1 of the columnar hollow portion 40 and the thickness t 3 of the small-diameter hollow portion whose opening diameter is narrowed by the reflecting surface 43 is the thickness t of the plate-like substrate 10.
  • the reflection surface 44 is a curved surface.
  • the pore 14 has an inner wall surface 14 a composed of an inner wall surface 41 of the columnar hollow portion 40 and a reflecting surface 44 formed of a curved surface.
  • the sum of the thickness t 1 of the columnar hollow portion 40 and the thickness t 4 in the pore length direction of the portion where the curved reflecting surface 44 is formed is the thickness t of the plate-like substrate 10. is there.
  • the reflecting surface 44 is a curved surface
  • the angle formed between the tangent at the position on the most surface 12 side of the reflecting surface 44 and the other surface 12 is ⁇ , and this angle ⁇ should be 0 ° to 45 °. That's fine. Also in this case, the same effect as the above embodiment can be obtained.
  • the inspection device of the third modification shown in FIG. 8 includes the same reflective surface 42 as that of the above-described embodiment. Furthermore, a tapered surface 45 whose opening diameter gradually increases from one columnar hollow portion 40 is provided in the pore opening portion on the one surface 11 side of the plate-like substrate 10.
  • the pore 14 has an inner wall surface 14 a composed of the inner wall surface 41, the reflecting surface 42, and the tapered surface 45 of the columnar hollow portion 40.
  • the sum of the thickness t 1 of the columnar hollow portion 40, the thickness t 2 occupied by the reflecting surface 42, and the thickness t 5 in the pore length direction of the portion where the tapered surface 45 is formed is a plate shape. It is the thickness t of the base material 10.
  • Increasing the opening diameter of the opening of one surface 11 as in this example can increase the amount of light extracted from the one surface 11 side of the light generated in the pores 14, which is preferable.
  • the inspection device of the design modification example 4 shown in FIG. 9 includes a reflective surface 42 on the other surface 12 side in succession to the inner wall surface 41 of the columnar hollow portion 40, as in the above-described embodiment.
  • a tapered surface 49 is provided on the other surface 12 side of 42 to increase the opening diameter again.
  • the pore 14 has an inner wall surface 14 a composed of the inner wall surface 41, the reflecting surface 42, and the tapered surface 49 of the columnar hollow portion 40.
  • the total of the thickness t 1 of the columnar hollow portion 40, the thickness t 2 occupied by the reflecting surface 42, and the thickness t 6 occupied by the tapered surface 49 is the thickness t of the plate-like substrate 10. In this way, by providing a portion having a larger opening diameter on the other surface 12 side than the reflecting surface 42, the inspection solution into the pores 14 from the other surface 12 side in the inspection method described later. The structure is easy to suck up.
  • the thickness of the columnar hollow 40 It is desirable that the ratio of t 1 to the opening diameter of the columnar hollow portion is 5 to 100.
  • the thickness t 1 of the columnar hollow portion 40 and the reflection are excluded except for the thickness t 6 occupied by the tapered surface 49. It is desirable that the ratio of the total t 1 + t 2 of the thickness t 2 occupied by the surface 42 to the opening diameter of the columnar hollow portion is 5 to 100.
  • the plate-like substrate 10 may be made of a single material, or may be made of a plurality of materials that differ depending on the part.
  • FIG. 10 shows a configuration example of an inspection device having portions made of different materials.
  • FIG. 10 is a cross-sectional view of a part of the inspection device.
  • the inspection device shown in FIG. 10 has pores 14 having the same shape as that of the inspection device of design modification example 3 shown in FIG.
  • the plate-like base material 10 includes a main body member 17 constituting the columnar hollow portion 40, and a first surface member 18 a constituting the tapered surface 45 and one surface 11 made of a material different from that of the main body member 17.
  • the second surface member 18b that constitutes the reflecting surface 42 and the other surface 12.
  • a plate-like main body member 17 made of Si, SiO 2 , Al, or Al 2 O 3 having a plurality of columnar pores is produced.
  • a main body member made of Si having a plurality of columnar pores can use a microporous Si manufacturing process known in MEMS (Micro Electro Mechanical System) technology.
  • the main body member made of SiO 2 having a plurality of columnar pores can be obtained by performing a thermal oxidation process on the main body member made of Si.
  • a plate-like body member made of Al having a plurality of columnar pores can be produced by forming pores in an Al plate-like substrate by a mechanical method such as a drill, or by etching or the like.
  • the production method of the main body member 17 is not limited to these, and various known techniques for forming a porous substrate can be used.
  • the opening diameter on one surface 11 of the plate-like base material 10 and the one surface 11 is made larger than that of the columnar hollow portion 40.
  • the first surface member 18a constituting the tapered surface 45 and the second surface member 18b constituting the reflecting surface 42 and the other surface 12 for reducing the opening diameter on the other surface 12 side of the columnar hollow portion 40 are: For example, it can be manufactured using a 3D printer.
  • a 3D printer is preferably a modeling method (powder sintering layered modeling method) in which a high-power laser beam is applied to a powdered material and sintered. In particular, the material powder and the laser are simultaneously irradiated to melt any part. If it is the thing of the modeling system made to make it laminate, the freedom degree of a foundation
  • the first material having a desired shape is formed by a metal material that can constitute a reflective surface, such as steel, stainless steel, aluminum, gold, platinum, silver, titanium, copper, nickel, cobalt, tin, tungsten, molybdenum, or niobium. And the 2nd surface member 18a, 18b can be produced easily.
  • the first and second surface members 18a and 18b made of a photocurable resin are produced by an optical modeling method in which a liquid photocurable resin composition is cured and laminated one by one with an ultraviolet laser. May be.
  • a reflective material may be selected as the photocurable resin.
  • UV (Ultraviolet) cured epoxy resin, UV cured acrylic resin, and the like can be given.
  • 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 to be used for inspection can be manufactured.
  • a known method can be applied without any particular limitation.
  • the method disclosed in the above-described Patent Document 1 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 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 catalyst of the reaction solution It is an optical signal resulting from a label, such as fluorescence or chemiluminescence.
  • a label such as fluorescence or chemiluminescence.
  • 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 solution is discharged, and light generated from a luminescence reaction that acts using the label F as a substrate is detected by the photodetector 70 in a state where the inspection solution such as a buffer solution is filled in the pores 14 (S4). ).
  • the inspection device 1 when the light emitted from the inspection device 1 is detected by the photodetector 70, the inspection device 1 includes the reflection surface 42, thereby reflecting the light generated in the pore 14 and emitted to the other surface 12 side. Since the light can be reflected to the one surface 11 side by the surface 42, light detection can be performed efficiently on the one surface 11 side.
  • 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 label that catalyzes a chemiluminescent substrate such as luminol
  • a reaction solution containing a chemiluminescent substrate that promotes a chemical reaction using the enzyme label as a catalyst after the washing treatment is used as a test solution.
  • 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 produce 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 reacting a luminol-based chemiluminescent substrate.
  • ALP alkaline phosphatase
  • 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.

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Abstract

La présente invention vise à fournir un dispositif d'inspection avec lequel il est possible d'améliorer l'efficacité d'extraction de lumière, en plus d'un appareil d'inspection équipé du dispositif d'inspection, et un procédé d'inspection. À cet effet, ce dispositif d'inspection (1) selon la présente invention comprend une pluralité de pores (14) qui traversent un substrat en forme de plaque (10) d'une première surface (11) à une seconde surface (12). Chacun des pores (14) a une cavité en colonne (40) qui s'étend dans le même diamètre d'ouverture, et une surface réfléchissante (42) qui est formée sur le côté de seconde surface (12) de la cavité en colonne (40), qui est contiguë à la surface de paroi interne de la cavité en colonne (40), et qui réduit le diamètre d'ouverture des pores à plus petit que le diamètre d'ouverture dans la cavité en colonne (40). L'angle formé par la surface réfléchissante (42) et la seconde surface (12) est compris entre 0° et 45°.
PCT/JP2017/010093 2016-03-30 2017-03-14 Dispositif d'inspection, appareil d'inspection et procédé d'inspection WO2017169715A1 (fr)

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JP2016-067480 2016-03-30

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Citations (7)

* 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
JP2004012206A (ja) * 2002-06-05 2004-01-15 Fuji Photo Film Co Ltd 測定チップ
JP2004191127A (ja) * 2002-12-10 2004-07-08 Nippon Sheet Glass Co Ltd 生化学容器
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
JP2005113136A (ja) * 2003-09-19 2005-04-28 Foundation For The Promotion Of Industrial Science 流体分散可能物質を基板上にパターニングする方法及び装置とそのための有孔シート材並びにその製造方法
JP4125244B2 (ja) * 2002-04-19 2008-07-30 インフィネオン テクノロジーズ アクチエンゲゼルシャフト 局部的に酸化された多孔質シリコンからなる装置、および、その製造方法
JP2009008672A (ja) * 2007-05-31 2009-01-15 Canon Inc 検知素子、検出装置、及び検知素子の製造方法

Patent Citations (7)

* 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
JP4125244B2 (ja) * 2002-04-19 2008-07-30 インフィネオン テクノロジーズ アクチエンゲゼルシャフト 局部的に酸化された多孔質シリコンからなる装置、および、その製造方法
JP2004012206A (ja) * 2002-06-05 2004-01-15 Fuji Photo Film Co Ltd 測定チップ
JP2004191127A (ja) * 2002-12-10 2004-07-08 Nippon Sheet Glass Co Ltd 生化学容器
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
JP2005113136A (ja) * 2003-09-19 2005-04-28 Foundation For The Promotion Of Industrial Science 流体分散可能物質を基板上にパターニングする方法及び装置とそのための有孔シート材並びにその製造方法
JP2009008672A (ja) * 2007-05-31 2009-01-15 Canon Inc 検知素子、検出装置、及び検知素子の製造方法

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