WO2016170967A1 - 検出チップの製造方法および検出チップ - Google Patents
検出チップの製造方法および検出チップ Download PDFInfo
- Publication number
- WO2016170967A1 WO2016170967A1 PCT/JP2016/061112 JP2016061112W WO2016170967A1 WO 2016170967 A1 WO2016170967 A1 WO 2016170967A1 JP 2016061112 W JP2016061112 W JP 2016061112W WO 2016170967 A1 WO2016170967 A1 WO 2016170967A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- hole
- frame
- detection chip
- support
- light
- Prior art date
Links
Images
Classifications
-
- 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
- G01N21/645—Specially adapted constructive features of fluorimeters
- G01N21/648—Specially adapted constructive features of fluorimeters using evanescent coupling or surface plasmon coupling for the excitation of fluorescence
-
- 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
- G01N21/01—Arrangements or apparatus for facilitating the optical investigation
- G01N21/03—Cuvette constructions
-
- 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
- G01N21/62—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
- G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
-
- 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
-
- 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
- G01N33/54366—Apparatus specially adapted for solid-phase testing
- G01N33/54373—Apparatus specially adapted for solid-phase testing involving physiochemical end-point determination, e.g. wave-guides, FETS, gratings
-
- 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
- G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
- G01N2021/6439—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" with indicators, stains, dyes, tags, labels, marks
-
- 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
- G01N21/645—Specially adapted constructive features of fluorimeters
- G01N2021/6482—Sample cells, cuvettes
-
- 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
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/55—Specular reflectivity
- G01N21/552—Attenuated total reflection
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2201/00—Features of devices classified in G01N21/00
- G01N2201/06—Illumination; Optics
- G01N2201/061—Sources
- G01N2201/06113—Coherent sources; lasers
- G01N2201/0612—Laser diodes
Definitions
- the present invention relates to a detection chip manufacturing method and a detection chip for detecting a substance to be detected.
- SPFS surface plasmon resonance fluorescence analysis
- a detection chip (sensor structure) having a sensor member and a well member fixed on the sensor member is used.
- the sensor member a prism (dielectric member), a metal film formed on the prism, and a capturing body (ligand) formed on the metal film for capturing a substance to be detected (analyte) are immobilized.
- a reaction field ligand immobilization region which is a defined region.
- the well member has a through hole at a position corresponding to the reaction field.
- a target substance is captured by a capturing body in a reaction field in a well, and the captured target substance is labeled with a fluorescent substance.
- the metal film is irradiated with excitation light through the prism at an angle at which surface plasmon resonance occurs, localized field light can be generated on the surface of the metal film.
- the fluorescent substance that labels the detection target substance captured on the metal film is selectively excited, and fluorescence is emitted from the fluorescent substance.
- the detection device can detect the presence or amount of the substance to be detected by detecting this fluorescence.
- the detection chip described in Patent Document 1 uses a frame member having a through-hole or a well member disposed on the metal film after the capture body is fixed on the entire surface of the metal film. After the capture body is fixed to a part of the surface of the metal film, the frame member is removed and a well member is newly disposed on the metal film. By manufacturing the detection chip in this way, it is possible to prevent the capturing body from being fixed to the inner wall surface of the through hole of the well member.
- An object of the present invention is to provide a detection chip manufacturing method capable of preventing the capture body from being fixed to the inner wall surface of the well without using a frame member that does not constitute the detection chip, and a detection chip manufactured by the manufacturing method. Is to provide.
- a method for manufacturing a detection chip is a method for manufacturing a detection chip used for detecting a substance to be detected in a specimen, and includes a first through hole.
- a detection chip is a detection chip used to detect a substance to be detected in a sample, and has a support and a first through hole.
- a capturing body for capturing the substance to be detected in the specimen is immobilized on the surface of the support exposed in the first through hole.
- the detection chip manufacturing method of the present invention it is possible to easily manufacture a detection chip in which the capturing body is not fixed to the inner wall surface of the well without using a frame member that does not constitute the detection chip.
- the detection chip obtained by the present invention it is possible to detect the substance to be detected with high sensitivity and high accuracy while suppressing loss of the substance to be detected due to adhesion to the inner wall surface of the well.
- FIG. 1 is a diagram showing a configuration of a surface plasmon enhanced fluorescence detection apparatus.
- 2A and 2B are diagrams showing the configuration of the detection chip according to the embodiment of the present invention.
- FIG. 3 is a flowchart showing an example of the manufacturing process of the detection chip according to the embodiment of the present invention.
- 4A to 4D are schematic views showing an example of the manufacturing process of the detection chip according to the embodiment of the present invention.
- FIG. 5 is a flowchart showing an example of an operation procedure of the surface plasmon enhanced fluorescence detection apparatus.
- 6A and 6B are diagrams illustrating a configuration of a detection chip according to the first modification.
- 7A and 7B are diagrams illustrating a configuration of a detection chip according to the second modification.
- SPFS apparatus surface plasmon enhanced fluorescence detection apparatus
- SPR surface plasmon resonance
- FIG. 1 is a diagram illustrating a configuration of the SPFS apparatus 100.
- the SPFS device 100 includes a chip holder 110 for detachably holding the detection chip 10, a light irradiation unit 120 for irradiating light to the detection chip 10, and emission from the detection chip 10.
- the SPFS device 100 is used with the detection chip 10 mounted on the chip holder 110. Therefore, the detection chip 10 will be described first, and then each component of the SPFS device 100 will be described.
- FIGS. 2A and 2B are diagrams showing the configuration of the detection chip 10 according to the present embodiment.
- 2A is a plan view of the detection chip 10
- FIG. 2B is a cross-sectional view taken along line BB in FIG. 2A.
- the detection chip 10 has an incident surface 21, a film formation surface 22, and an emission surface 23, and a prism (support) 20 having a metal film 30 formed on the film formation surface 22.
- a first frame 40 disposed on the metal film 30 and having the first through-hole 41, and a second frame body disposed on the metal film 30 in the first through-hole 41 and having the second through-hole 51. 50.
- the second frame 50 is disposed on the prism 20 so as to close one opening of the second through hole 51, whereby a well 53 for containing a liquid is formed.
- the prism 20 is made of a dielectric that is transparent to the excitation light ⁇ . As described above, the prism 20 has the incident surface 21, the film formation surface 22, and the emission surface 23.
- the incident surface 21 causes the excitation light ⁇ from the light irradiation unit 120 to enter the prism 20.
- a metal film 30 is disposed on the film formation surface 22.
- the excitation light ⁇ incident on the inside of the prism 20 is reflected by the metal film 30. More specifically, the excitation light ⁇ is reflected at the interface (deposition surface 22) between the prism 20 and the metal film 30.
- the emission surface 23 emits the excitation light ⁇ reflected by the metal film 30 to the outside of the prism 20.
- the shape of the prism 20 is not particularly limited.
- the shape of the prism 20 is a column having a trapezoidal bottom surface.
- the surface corresponding to one base of the trapezoid is the film formation surface 22, the surface corresponding to one leg is the incident surface 21, and the surface corresponding to the other leg is the emission surface 23.
- the trapezoid serving as the bottom surface is preferably an isosceles trapezoid.
- the entrance surface 21 and the exit surface 23 are symmetric, and the S wave component of the excitation light ⁇ is less likely to stay in the prism 20.
- the incident surface 21 is formed so that the excitation light ⁇ does not return to the light irradiation unit 120.
- the light source of the excitation light ⁇ is a laser diode (hereinafter also referred to as “LD”)
- LD laser diode
- the excitation state of the LD is disturbed, and the wavelength and output of the excitation light ⁇ vary. Because it ends up. Therefore, the angle of the incident surface 21 is set so that the excitation light ⁇ does not enter the incident surface 21 perpendicularly in a scanning range centered on an ideal resonance angle or enhancement angle.
- the “resonance angle” refers to the incident angle when the amount of reflected light (not shown) emitted from the emission surface 23 is minimum when the incident angle of the excitation light ⁇ with respect to the metal film 30 is scanned.
- the “enhancement angle” refers to scattered light having the same wavelength as the excitation light ⁇ emitted above the detection chip 10 when the incident angle of the excitation light ⁇ with respect to the metal film 30 is scanned (hereinafter referred to as “plasmon scattered light”). This means the angle of incidence when the light quantity of ⁇ is maximized.
- the angle between the incident surface 21 and the film formation surface 22 and the angle between the film formation surface 22 and the emission surface 23 are both about 80 °.
- the prism 20 may have other elements as necessary.
- the prism 20 may have a reagent holder for storing the reagent at a position that does not hinder the optical path of the excitation light ⁇ .
- Examples of the material of the prism 20 include resin and glass.
- Examples of the resin constituting the prism 20 include polymethyl methacrylate (PMMA), polycarbonate (PC), and cycloolefin-based polymer.
- the prism 20 is preferably a resin having a refractive index of 1.4 to 1.6 and a small birefringence.
- the metal film 30 is formed on one surface (deposition surface 22) of the prism 20.
- an interaction surface plasmon resonance; SPR
- SPR surface plasmon resonance
- the material of the metal film 30 is not particularly limited as long as it is a metal that causes surface plasmon resonance.
- Examples of the material of the metal film 30 include gold, silver, copper, aluminum, and alloys thereof.
- the metal film 30 is a gold thin film.
- the thickness of the metal film 30 is not particularly limited, but is preferably in the range of 30 to 70 nm.
- a capture body 60 for capturing a substance to be detected is fixed to the surface of the metal film 30 that does not face the prism 20. More specifically, the capturing body 60 is fixed to at least a part of the inner wall surface of the first through hole 41 of the first frame body 40 and the surface of the metal film 30 exposed in the first through hole 41. (See FIG. 4C described later).
- 2B in order to illustrate the second frame body 50, the capturing body 60 fixed to the inner wall surface of the first through-hole 41 and the second frame body 50 among the surfaces of the metal film 30 are arranged. The capturing body 60 fixed to the portion that is formed is omitted. The capturing body 60 can selectively detect the substance to be detected.
- At least a part of the surface of the metal film 30 on which the capturing body 60 is immobilized undergoes a reaction such as binding between the capturing body 60 and the substance to be detected (primary reaction) or a fluorescent label (secondary reaction) of the substance to be detected. It is set as the reaction field to be performed.
- the surface of the metal film 30 exposed in the second through hole 51 of the second frame 50 described later is set as the reaction field.
- the type of the capturing body 60 is not particularly limited as long as the target substance can be captured.
- the capture body 60 is an antibody or a fragment thereof that can specifically bind to the substance to be detected.
- the first frame body 40 has a first through hole 41 and is disposed on the prism 20 (metal film 30) so that one opening of the first through hole 41 is closed.
- the first frame body 40 defines a region on the metal film 30 where the capturing body 60 is to be immobilized when the detection chip 10 according to the present embodiment is manufactured.
- the number, shape, and size of the first through holes 41 are not particularly limited, and can be set as appropriate according to the application of the detection chip 10. In the present embodiment, the number of the first through holes 41 is one, and the shape of the first through holes 41 is a cylindrical shape.
- the first frame 40 preferably has a light shielding property. As a result, noise light such as autofluorescence and external light emitted from the prism 20 can be cut, and the noise light can be prevented from reaching the light receiving unit 130 of the SPFS device 100.
- the outer shape of the first frame body 40 is not particularly limited.
- the planar view shape of the outer shape of the first frame body 40 is a circular shape, a square shape, or the like.
- the planar view shape of the outer shape of the first frame body 40 is a quadrangular shape.
- the first frame 40 may have other elements as necessary.
- the first frame body 40 may have a reagent holder 43 for containing the reagent.
- the reagent holder 43 may be integrated with the first frame body 40 or may be a separate body. In the present embodiment, the reagent holder 43 is integrated with the first frame body 40.
- the material of the first frame body 40 examples include resin and glass.
- the first frame 40 may be a resin film. Thereby, manufacture becomes easy, manufacturing cost can be reduced, and the detection chip 10 can be miniaturized.
- the second frame 50 has a second through hole 51, and on the prism 20 (metal film 30) in the first through hole 41 so that one opening of the second through hole 51 is closed. Has been placed.
- the second frame 50 may be disposed so as to be in contact with the first frame 40 without a gap, or may be disposed separately.
- the second frame 50 defines a well 53 for containing a liquid.
- the second frame 50 serves as a mark, and the irradiation position of the excitation light ⁇ in the SPFS device 100 It becomes easy to adjust the position.
- the shape and size of the second through hole 51 are not particularly limited, and can be set as appropriate according to the application.
- Examples of the shape of the second through hole 51 include a cylindrical trapezoidal shape, a cylindrical shape, an elliptical column shape, a polygonal column shape, and combinations thereof.
- the shape of the second through hole 51 is preferably a columnar shape or an elliptical column shape.
- the shape of the second through hole 51 is a combination of a columnar shape and a columnar trapezoidal shape.
- the depth h2 of the second through hole 51 is preferably higher than the height h1 of the first through hole 41. Thereby, it is possible to prevent the liquid from splashing out of the well 53 when the liquid is provided in the well 53.
- the height of the second through-hole 51 can be appropriately set according to the height or amount of the liquid metal film 30 to be provided and the amount of rebound from the inner wall surface of the second through-hole 51.
- the outer shape of the second frame 50 is not particularly limited.
- the planar view shape of the outer shape of the second frame 50 is a circular shape or a quadrangular shape.
- the plan view shape of the outer shape of the second frame 50 is a circular shape.
- the second frame 50 preferably has a tapered portion in which the cross-sectional area of the second through hole 51 in the direction orthogonal to the height direction of the second through hole 51 increases as the distance from the prism 20 increases. Thereby, it becomes easy to provide a liquid in the 2nd through-hole 51 (well 53).
- the second frame 50 has a tapered portion 52.
- the inner wall surface of the tapered portion 52 is emitted from the center of the surface (reaction field) of the metal film 30 exposed in the second through hole 51 and is within 20 °. It is more preferable to incline so as not to block the light beam emitted at the emission angle. Thereby, it is possible to suppress the fluorescence ⁇ released from the fluorescent substance that labels the substance to be detected from being blocked, and to detect the substance to be detected with high accuracy.
- the inner wall surface of the second through hole 51 is preferably subjected to a blocking process. Thereby, the nonspecific binding of the substance to be detected to the inner wall surface of the second through hole 51 can be suppressed, and the detection efficiency of the substance to be detected can be improved.
- the second frame 50 may have other elements as necessary.
- the second frame 50 may have a reagent holder for containing the reagent.
- the shape and color of the inner wall surface of the second frame 50 can be set as appropriate.
- the inner wall surface of the second frame 50 may be black from the viewpoint of absorbing excess noise light.
- minute unevenness may be provided on the inner wall surface of the second frame 50 from the viewpoint of reducing noise by dispersing the noise light.
- the color of the inner wall surface of the second frame 50 may be white, or the second frame.
- the inner wall surface of 50 may be a mirror surface.
- the material of the second frame 50 examples include resin and glass.
- the second frame 50 may be a resin film. Thereby, manufacture becomes easy, a manufacturing cost can be reduced, and the detection chip 10 can also be reduced in size.
- the second frame 50 is bonded to the metal film 30 or the prism 20 by, for example, adhesion using a double-sided tape or an adhesive, laser welding, ultrasonic welding, or pressure bonding using a clamp member. Further, the second frame body 50 may be joined to the first frame body 40.
- the excitation light ⁇ guided to the prism 20 enters the prism 20 through the incident surface 21.
- the excitation light ⁇ incident on the prism 20 is incident on the interface (deposition surface 22) between the prism 20 and the metal film 30 so as to have a total reflection angle (an angle at which surface plasmon resonance occurs).
- the reflected light reflected at the interface is emitted outside the prism 20 at the emission surface 23 (not shown).
- plasmon scattered light ⁇ , fluorescence ⁇ from a fluorescent substance, and the like are emitted from the reaction field above the detection chip 10.
- FIG. 3 is a flowchart showing an example of the manufacturing process of the detection chip 10 according to the present embodiment.
- 4A to 4D are schematic views showing an example of the manufacturing process of the detection chip 10 according to the present embodiment.
- the metal film 30 is formed on one surface (film formation surface 22) having an incident surface 21, a film formation surface 22, and an emission surface 23.
- a prism 20 (support) is prepared (step S10).
- the prism 20 is formed into a desired shape.
- the method for forming the prism 20 is not particularly limited.
- the prism 20 may be formed by a mold forming method.
- a metal film 30 is formed on the film formation surface 22 of the prism 20. Examples of the method for forming the metal film 30 include sputtering, vapor deposition, and plating. Moreover, you may purchase the prism 20 in which the metal film 30 was already formed.
- the first frame 40 having the first through-hole 41 is replaced with the metal film 30 so that one opening of the first through-hole 41 is closed. It arrange
- the method of forming the first through hole 41 in the first frame body 40 is not particularly limited.
- the first through hole 41 may be formed by a mold forming method, a cutting process, or the like.
- step S30 the capturing body 60 is fixed on the metal film 30 exposed in the first through hole 41 (step S30). Specifically, a liquid including the capturing body 60 is provided in the first through hole 41 of the first frame body 40 disposed on the metal film 30. Accordingly, the capturing body 60 is formed on at least a part of the inner wall surface of the first through hole 41 of the first frame body 40 that is in contact with the provided liquid and on the surface of the metal film 30 exposed in the first through hole 41. Can be immobilized.
- the method for immobilizing the capturing body 60 on the metal film 30 is not particularly limited.
- a self-assembled monomolecular film hereinafter referred to as “SAM”
- SAMs include films formed with substituted aliphatic thiols such as HOOC— (CH 2 ) 11 —SH.
- the material constituting the polymer film include polyethylene glycol and MPC polymer.
- a polymer having a reactive group that can be bound to the capturing body 60 (or a functional group that can be converted into a reactive group) may be fixed to the metal film 30 and the capturing body 60 may be bound to the polymer.
- the fourth step after removing the liquid including the capturing body 60 from the metal film 30, the second frame body 50 having the second through-hole 51 is replaced with the second frame 50 as shown in FIG. 4D. It arrange
- the second through hole 51 can be formed by a method similar to that of the first through hole 41, for example.
- the detection chip 10 used for detecting the substance to be detected contained in the specimen can be manufactured.
- a step of blocking the second frame 50 may be further included.
- a liquid containing a blocking agent may be brought into contact with the inner wall surface of the second frame 50.
- the blocking process may be performed in advance before the second frame body 50 is disposed on the metal film 30, or may be performed after the second frame body 50 is disposed on the metal film 30.
- the blocking agent include casein, skim milk, albumin (including bovine serum albumin), gelatin, high molecular compounds such as polyethylene glycol; phospholipid, low molecular weight compounds such as ethylenediamine and acetonitrile, and the like. These may be used alone or in combination of two or more.
- an optional step may include a step of protecting the capturing body 60 with a moisturizing agent in order to prevent the capturing body 60 from drying.
- the capturing body 60 is also fixed to a part of the inner wall surface of the first through hole 41 of the first frame body 40.
- the second frame 50 disposed inside the first through hole 41 defines the well 53, the liquid introduced into the well 53 does not contact the inner wall surface of the first through hole 41.
- the capturing body 60 is not fixed to the inner wall surface of the second through hole 51 of the second frame 50. For this reason, when the specimen is introduced into the well 53, the substance to be detected contained in the specimen is efficiently captured by the capturing body 60 immobilized on the bottom surface of the well 53.
- the SPFS device 100 includes the chip holder 110, the light irradiation unit 120, the light receiving unit 130, and the control unit 140 (see FIG. 1).
- the chip holder 110 holds the detection chip 10 at a predetermined position.
- the detection chip 10 is irradiated with the excitation light ⁇ from the light irradiation unit 120 while being held by the chip holder 110.
- the light irradiation unit 120 irradiates excitation light ⁇ (single mode laser light) toward the incident surface 21 of the prism 20 of the detection chip 10 held by the chip holder 110. More specifically, the light source unit 121 emits the excitation light ⁇ to the region corresponding to the well 53 in the back surface of the metal film 30 so as to have a total reflection angle.
- the light irradiation unit 120 includes a light source unit 121 that emits excitation light ⁇ , an angle adjustment unit 122 that adjusts the incident angle of the excitation light ⁇ with respect to the interface (deposition surface 22) between the prism 20 and the metal film 30, and a light source unit. And a light source control unit 123 that controls various devices included in the device 121.
- the light source unit 121 emits excitation light ⁇ .
- the light source unit 121 includes a light source of excitation light ⁇ , a beam shaping optical system, an APC mechanism, and a temperature adjustment mechanism (all not shown).
- the type of light source is not particularly limited. Examples of light source types include laser diodes (LDs), light emitting diodes, mercury lamps, and other laser light sources.
- LDs laser diodes
- LEDs light emitting diodes
- mercury lamps mercury lamps
- other laser light sources other laser light sources.
- the excitation light ⁇ emitted from the light source is converted into a beam by a lens, a mirror, a slit, or the like.
- the excitation light ⁇ emitted from the light source is converted into monochromatic light by a diffraction grating or the like.
- the excitation light ⁇ emitted from the light source is not linearly polarized light, the excitation light ⁇ emitted from the light source is converted into linearly polarized light by a polarizer or the like.
- the beam shaping optical system includes, for example, a collimator, a band pass filter, a linear polarization filter, a half-wave plate, a slit, and a zoom means.
- the beam shaping optical system may include all of these or a part thereof.
- the collimator collimates the excitation light ⁇ emitted from the light source.
- the bandpass filter turns the excitation light ⁇ emitted from the light source into a narrow band light having only the center wavelength. This is because the excitation light ⁇ from the light source has a slight wavelength distribution width.
- the linear polarization filter turns the excitation light ⁇ emitted from the light source into completely linearly polarized light.
- the half-wave plate adjusts the polarization direction of the excitation light ⁇ so that the P-wave component light is incident on the metal film 30.
- the slit and zoom means adjust the beam diameter, contour shape, and the like of the excitation light ⁇ so that the shape of the irradiation spot on the back surface of the metal film 30 is a circle of a predetermined size.
- the APC mechanism controls the light source so that the output of the light source is constant. More specifically, the APC mechanism detects the amount of light branched from the excitation light ⁇ with a photodiode (not shown) or the like. The APC mechanism controls the input energy by a regression circuit, thereby controlling the output of the light source to be constant.
- the temperature adjustment mechanism is, for example, a heater or a Peltier element.
- the wavelength and energy of the light emitted from the light source may vary depending on the temperature. For this reason, the wavelength and energy of the light emitted from the light source are controlled to be constant by keeping the temperature of the light source constant by the temperature adjusting mechanism.
- the angle adjusting unit 122 adjusts the incident angle of the excitation light ⁇ to the metal film 30 (the interface between the prism 20 and the metal film 30 (film formation surface 22)).
- the angle adjusting unit 122 relatively rotates the optical axis of the excitation light ⁇ and the chip holder 110 in order to irradiate the excitation light ⁇ to a predetermined position of the metal film 30 (deposition surface 22) at a predetermined incident angle.
- the angle adjustment unit 122 rotates the light source unit 121 on the metal film 30 with an axis orthogonal to the optical axis of the excitation light ⁇ (an axis perpendicular to the paper surface of FIG. 1) as the central axis.
- the light source control unit 123 controls various devices included in the light source unit 121 to adjust the power of the excitation light ⁇ from the light source unit 121, the irradiation time, and the like.
- the light source control unit 123 includes, for example, a known computer or microcomputer including an arithmetic device, a control device, a storage device, an input device, and an output device.
- the light receiving unit 130 is disposed so as to face the surface of the metal film 30 of the detection chip 10 held by the chip holder 110 that does not face the prism 20.
- the light receiving unit 130 detects light (plasmon scattered light ⁇ or fluorescence ⁇ ) emitted from the metal film 30 in the second through hole 51.
- the light receiving unit 130 includes a first lens 132, an optical filter 133, a second lens 134, and a light receiving sensor 135 disposed in the light receiving optical system unit 131, a position switching mechanism 136, and an optical sensor control unit 137.
- the first lens 132 is, for example, a condensing lens, and condenses light emitted from the metal film 30.
- the second lens 134 is, for example, an imaging lens, and forms an image of the light collected by the first lens 132 on the light receiving surface of the light receiving sensor 135. The optical path between both lenses is substantially parallel.
- the optical filter 133 is disposed between the first lens 132 and the second lens 134.
- the optical filter 133 transmits only the fluorescence component of the incident light and removes the excitation light component (plasmon scattered light ⁇ ).
- the optical filter 133 removes the excitation light component, the fluorescence ⁇ can be detected with a high S / N ratio.
- types of the optical filter 133 include an excitation light reflection filter, a short wavelength cut filter, and a band pass filter.
- the light receiving sensor 135 detects plasmon scattered light ⁇ and fluorescence ⁇ emitted from the detection chip 10.
- the type of the light receiving sensor 135 is not particularly limited as long as the above object can be achieved. However, it is preferable that the variation in the detected value is small even when the amount of received light increases.
- the light receiving sensor 135 is, for example, a photodiode (PD).
- the position switching mechanism 136 switches the position of the optical filter 133 on or off the optical path in the light receiving optical system unit 131. Specifically, when measuring the optical blank value or the fluorescence value, the optical filter 133 is disposed on the optical path in the light receiving optical system unit 131, and when the light receiving sensor 135 detects the plasmon scattered light ⁇ , the optical filter 133 is placed in the optical path. Place outside.
- the light sensor control unit 137 detects the output value of the light receiving sensor 135, manages the sensitivity of the light receiving sensor 135 based on the detected output value, and controls the sensitivity of the light receiving sensor 135 to obtain an appropriate output value.
- the optical sensor control unit 137 includes, for example, a known computer or microcomputer including an arithmetic device, a control device, a storage device, an input device, and an output device.
- the control unit 140 controls the angle adjustment unit 122, the light source control unit 123, the position switching mechanism 136, and the optical sensor control unit 137.
- the control unit 140 also functions as a processing unit for calculating a signal value indicating the presence or amount of the substance to be detected based on the detection result of the light receiving sensor 135.
- the control unit 140 includes, for example, a known computer or microcomputer including an arithmetic device, a control device, a storage device, an input device, and an output device.
- FIG. 5 is a flowchart illustrating an example of an operation procedure of the SPFS apparatus.
- step S110 preparation for detection is performed (step S110). Specifically, the detection chip 10 is installed in the chip holder 110 of the SPFS device 100. Further, when a humectant is present in the well 53 of the detection chip 10, the well 53 is washed to remove the humectant so that the capturing body 60 can appropriately capture the substance to be detected.
- the enhancement angle is determined (step S120). Specifically, the irradiation angle of the excitation light ⁇ with respect to the metal film 30 (deposition surface 22) is scanned while irradiating the predetermined position of the metal film 30 (deposition surface 22) with the excitation light ⁇ , and the optimum Determine the angle of incidence.
- the control unit 140 controls the light source control unit 123 and the angle adjustment unit 122 to irradiate the excitation light ⁇ from the light source unit 121 to a predetermined position of the metal film 30 (deposition surface 22), while forming the metal film 30 (the formation film).
- the incident angle of the excitation light ⁇ with respect to the film surface 22) is scanned.
- control unit 140 controls the position switching mechanism 136 to move the optical filter 133 out of the optical path of the light receiving optical system unit 131.
- control unit 140 controls the optical sensor control unit 137 so that the light receiving sensor 135 detects the plasmon scattered light ⁇ .
- the control unit 140 obtains data including the relationship between the incident angle of the excitation light ⁇ and the intensity of the plasmon scattered light ⁇ . Then, the control unit 140 analyzes the data and determines an incident angle (enhancement angle) that maximizes the intensity of the plasmon scattered light ⁇ .
- the enhancement angle is determined by the material and shape of the prism 20, the thickness of the metal film 30, the refractive index of the liquid in the well 53, etc., but depends on various factors such as the type and amount of the capturing body 60 and the shape error of the prism 20. Slightly fluctuates. For this reason, it is preferable to determine the enhancement angle each time detection is performed. The enhancement angle is determined on the order of about 0.1 °.
- the incident angle of the excitation light ⁇ with respect to the metal film 30 (deposition surface 22) is set to the enhancement angle determined in step S120 (step S130). Specifically, the control unit 140 controls the angle adjustment unit 122 to set the incident angle of the excitation light ⁇ with respect to the metal film 30 (deposition surface 22) as an enhancement angle. In the subsequent steps, the incident angle of the excitation light ⁇ with respect to the metal film 30 (deposition surface 22) remains the enhancement angle.
- the optical blank value means the amount of background light emitted above the detection chip 10. This background light is mainly caused by autofluorescence or external light emitted from the detection chip 10 (prism 20) when the excitation light ⁇ is irradiated.
- control unit 140 controls the position switching mechanism 136 to move the optical filter 133 onto the optical path of the light receiving optical system unit 131.
- control unit 140 controls the light source control unit 123 to transmit the metal from the light source unit 121 through the prism 20 so that surface plasmon resonance occurs in the metal film 30 in the absence of the fluorescent material on the metal film 30.
- the film 30 is irradiated with excitation light ⁇ .
- control unit 140 controls the light sensor control unit 137 to detect the light emitted from the detection chip 10 by the light receiving sensor 135 to obtain an optical blank value.
- the measured optical blank value is transmitted to the control unit (processing unit) 140 and stored.
- the substance to be detected in the specimen and the capturing body 60 are reacted (primary reaction; step S150). Specifically, the specimen is injected into the well 53, and the specimen and the capturing body 60 are brought into contact with each other. When the substance to be detected exists in the specimen, at least a part of the substance to be detected is captured by the capturing body 60. At this time, the substance to be detected is appropriately captured by the capturing body 60 immobilized on the bottom surface of the well 53. The capture body 60 is also immobilized on a part of the inner wall surface of the first through hole 41 of the first frame body 40, but contact between the capture body 60 immobilized on a portion other than the bottom surface of the well 53 and the specimen is not performed. This is hindered by the second frame 50.
- the capturing body 60 is not fixed to the inner wall surface of the second through hole 51 of the second frame 50. For this reason, the substance to be detected is efficiently captured by the capturing body 60 immobilized on the bottom surface of the well 53. Thereafter, the inside of the well 53 is washed with a buffer solution or the like to remove substances not captured by the capturing body 60.
- the type of specimen is not particularly limited. Examples of the specimen include body fluids such as blood, serum, plasma, urine, nasal fluid, saliva, semen, and diluted solutions thereof.
- the target substance captured by the capturing body 60 is labeled with a fluorescent substance (secondary reaction; step S160).
- a fluorescent labeling solution is provided in the well 53.
- the fluorescent labeling solution is, for example, a buffer solution containing an antibody (secondary antibody) labeled with a fluorescent substance.
- the fluorescent labeling solution comes into contact with the target substance captured by the capturing body 60, and the target substance is labeled with the fluorescent substance. Thereafter, the inside of the well 53 is washed with a buffer solution or the like to remove free fluorescent substances.
- the excitation light ⁇ is irradiated onto the metal film 30 (film formation surface 22), and the fluorescent substance labeling the detection target substance in the reaction field is started.
- the emitted fluorescence ⁇ is detected and the fluorescence value is measured (step S170).
- the control unit 140 controls the light source control unit 123 so that the surface plasmon resonance is generated in the metal film 30 in a state where the detection target substance labeled with the fluorescent substance exists on the metal film 30.
- the excitation light ⁇ is emitted from the light source unit 121 to the metal film 30 through the prism 20.
- control unit 140 controls the optical sensor control unit 137 to detect the fluorescence ⁇ emitted from the fluorescent substance that labels the substance to be detected by the light receiving sensor 135.
- the measured fluorescence value is transmitted to the control unit (processing unit) 140 and stored.
- a signal value indicating the presence or amount of the substance to be detected is calculated (step S180). Specifically, the control unit 140 calculates a signal value correlated with the amount of the substance to be detected by subtracting the optical blank value obtained in step S140 from the fluorescence value obtained in step S170. The signal value can be converted into the amount and concentration of the substance to be detected by a calibration curve prepared in advance.
- the detection target substance can be detected with high accuracy using the detection chip 10 according to the present embodiment.
- step S150 After performing the primary reaction (step S150), the determination of the enhancement angle (step S120), the setting of the incident angle to the enhancement angle (step S130), and the measurement of the optical blank value (step S140) may be performed. .
- FIGS. 6A and 6B are diagrams illustrating the configuration of the detection chip 10 ′ according to the first modification.
- 6A is a plan view of the detection chip 10 '
- FIG. 6B is a cross-sectional view taken along line BB in FIG. 6A.
- the first frame body 40 ′ increases in cross-sectional area of the first through hole 41 ′ in the direction orthogonal to the height direction of the first through hole 41 ′ as it is away from the prism 20.
- the second frame 50 ′ may be disposed so as to be in contact with the first frame 40 ′ without a gap or may be disposed apart from the first frame 40 ′.
- the shape of the second frame 50 ′ is formed so as to be in contact with the first frame 40 ′ without a gap.
- FIGS. 7A and 7B are diagrams showing the configuration of the detection chip 10 ′′ according to the modification 2.
- FIG. 7A is a plan view of the detection chip 10 ′′
- FIG. 7B is a cross-sectional view taken along line BB in FIG. 7A.
- the shape of the well 53 ′′ is a cylindrical shape. Also in this case, the same effects as those in the above embodiment can be obtained by manufacturing the detection chip 10 ′′ in the same manner as in the above embodiment.
- the detection chip according to the present invention includes the detection chip 10, 10 ′, 10 ′′.
- a support such as a glass substrate may be used instead of the prism 20 having the metal film 30 formed on one surface.
- the detection chip according to the present invention can detect a substance to be detected with high sensitivity and high accuracy, and is useful for clinical examinations, for example.
Landscapes
- Health & Medical Sciences (AREA)
- Immunology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Physics & Mathematics (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Analytical Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Hematology (AREA)
- Molecular Biology (AREA)
- Urology & Nephrology (AREA)
- Biomedical Technology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Cell Biology (AREA)
- Medicinal Chemistry (AREA)
- Food Science & Technology (AREA)
- Microbiology (AREA)
- Biotechnology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Optics & Photonics (AREA)
- Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
- Optical Measuring Cells (AREA)
Abstract
Description
図2A、Bは、本実施の形態に係る検出チップ10の構成を示す図である。図2Aは、検出チップ10の平面図であり、図2Bは、図2AにおけるB-B線の断面図である。図2A、Bに示されるように、検出チップ10は、入射面21、成膜面22および出射面23を有し、成膜面22上に金属膜30が形成されたプリズム(支持体)20と、金属膜30上に配置され、第1貫通孔41を有する第1枠体40と、金属膜30上において第1貫通孔41内に配置され、第2貫通孔51を有する第2枠体50とを有する。第2枠体50が第2貫通孔51の一方の開口を閉塞されるようにプリズム20上に配置されることにより、液体を収容するためのウェル53が形成される。
次に、本実施の形態に係る検出チップ10の製造方法の一例について説明する。図3は、本実施の形態に係る検出チップ10の製造工程の一例を示すフローチャートである。図4A~Dは、本実施の形態に係る検出チップ10の製造工程の一例を示す模式図である。
第1工程では、図4Aに示されるように、入射面21、成膜面22および出射面23を有し、一面(成膜面22)上に金属膜30が形成されたプリズム20(支持体)を準備する(工程S10)。まず、プリズム20を所望の形状に成形する。プリズム20を成形する方法は、特に限定されず、例えば、金型成形法によりプリズム20を成形すればよい。次いで、プリズム20の成膜面22上に金属膜30を形成する。金属膜30の形成方法の例には、スパッタリング、蒸着およびメッキが含まれる。また、金属膜30が既に形成されたプリズム20を購入してもよい。
第2工程では、図4Bに示されるように、第1貫通孔41を有する第1枠体40を、第1貫通孔41の一方の開口が閉塞されるように金属膜30上に配置する(工程S20)。また、第1枠体40に第1貫通孔41を形成する方法は、特に限定されず、例えば、金型成形法や切削加工などにより第1貫通孔41を形成すればよい。
第3工程では、図4Cに示されるように、第1貫通孔41内に露出した金属膜30上に捕捉体60を固定化する(工程S30)。具体的には、金属膜30上に配置された第1枠体40の第1貫通孔41内に捕捉体60を含む液体を提供する。これにより、提供された液体に接触した第1枠体40の第1貫通孔41の内壁面の少なくとも一部と、第1貫通孔41内に露出した金属膜30の表面とに、捕捉体60を固定化することができる。
第4工程では、金属膜30上から捕捉体60を含む液体を除去した後、図4Dに示されるように、第2貫通孔51を有する第2枠体50を、第2貫通孔51の一方の開口が閉塞されるように第1貫通孔41内の金属膜30上に配置する(工程S40)。通常は、捕捉体60を含む液体を除去した後、第2枠体50を配置する前に、緩衝液などで第1貫通孔41内を洗浄する。なお、図4Dには示されていないが、第2枠体50を第1貫通孔41内に配置しても、第1枠体40の第1貫通孔41の内壁面の一部と、金属膜30の表面とには、捕捉体60が固定化されている。第2貫通孔51は、例えば、第1貫通孔41と同様の方法により形成されうる。
次に、SPFS装置100の各構成要素について説明する。前述のとおり、SPFS装置100は、チップホルダー110、光照射ユニット120、受光ユニット130および制御部140を有する(図1参照)。
次に、検出チップ10を使用したSPFS装置100の動作について説明する。図5は、SPFS装置の動作手順の一例を示すフローチャートである。
以上のように、本実施の形態に係る検出チップ10を使用すれば、ウェル53の内壁面への被検出物質の付着を抑制して、検出されない被検出物質の数を低減させることができる。このため、検体中の被検出物質を高精度に検出することができる。
20 プリズム
21 入射面
22 成膜面
23 出射面
30 金属膜
40、40’ 第1枠体
41、41’ 第1貫通孔
50、50’、50” 第2枠体
51、51’、51” 第2貫通孔
42’、52 テーパー部
43 試薬ホルダー
53、53” ウェル
60 捕捉体
100 表面プラズモン共鳴蛍光分析装置(SPFS装置)
110 チップホルダー
120 光照射ユニット
121 光源ユニット
122 角度調整部
123 光源制御部
130 受光ユニット
131 受光光学系ユニット
132 第1レンズ
133 光学フィルター
134 第2レンズ
135 受光センサー
136 位置切替え機構
137 光センサー制御部
140 制御部(処理部)
α 励起光
β プラズモン散乱光
γ 蛍光
h1 第1貫通孔の高さ
h2 第2貫通孔の高さ
Claims (20)
- 検体中の被検出物質を検出するために使用される検出チップの製造方法であって、
第1貫通孔を有する第1枠体を、前記第1貫通孔の一方の開口が閉塞されるように支持体上に配置する工程と、
前記第1貫通孔内に被検出物質を捕捉するための捕捉体を含む液体を提供し、前記第1貫通孔内に露出した前記支持体上に前記捕捉体を固定化する工程と、
前記支持体上から前記液体を除去した後、第2貫通孔を有する第2枠体を、前記第2貫通孔の一方の開口が閉塞されるように前記第1貫通孔内の前記支持体上に配置する工程と、
を含む、検出チップの製造方法。 - 前記第1枠体は、遮光性を有する、請求項1に記載の検出チップの製造方法。
- 前記第1枠体は、前記支持体から離れるにつれて前記第1貫通孔の高さ方向に直交する方向における前記第1貫通孔の断面積が大きくなるテーパー部を有する、請求項1または請求項2に記載の検出チップの製造方法。
- 前記第2枠体は、前記支持体から離れるにつれて前記第2貫通孔の高さ方向に直交する方向における前記第2貫通孔の断面積が大きくなるテーパー部を有する、請求項1~3のいずれか一項に記載の検出チップの製造方法。
- 前記第2貫通孔の高さは、前記第1貫通孔の高さより高い、請求項1~4のいずれか一項に記載の検出チップの製造方法。
- 前記第2枠体の前記テーパー部の内壁面は、前記第2貫通孔内に露出した前記支持体の表面の中心から出射され、かつ20°以内の出射角で出射された光線を遮断しないように傾斜している、請求項4に記載の検出チップの製造方法。
- 前記第2貫通孔の内壁面をブロッキング処理する工程をさらに含む、請求項1~6のいずれか一項に記載の検出チップの製造方法。
- 前記支持体、前記第1枠体および前記第2枠体のうち少なくとも1つは、試薬を収容するための試薬ホルダーを有する、請求項1~7のいずれか一項に記載の検出チップの製造方法。
- 前記第1枠体および前記第2枠体のうち少なくとも1つは、樹脂フィルムからなる、請求項1~8のいずれか一項に記載の検出チップの製造方法。
- 前記支持体は、一面上に金属膜が形成された、誘電体からなるプリズムであり、
前記第1枠体および前記第2枠体は、前記一面上に配置される、
請求項1~9のいずれか一項に記載の検出チップの製造方法。 - 検体中の被検出物質を検出するために使用される検出チップであって、
支持体と、
第1貫通孔を有し、前記第1貫通孔の一方の開口が閉塞されるように前記支持体上に配置されている第1枠体と、
第2貫通孔を有し、前記第2貫通孔の一方の開口が閉塞されるように前記第1貫通孔内の前記支持体上に配置されている第2枠体と、
を有し、
前記第1枠体の前記第1貫通孔の内壁面の少なくとも一部と、前記第1貫通孔に露出した前記支持体の表面とに、検体中の被検出物質を捕捉するための捕捉体が固定化されている、
検出チップ。 - 前記第1枠体は、遮光性を有する、請求項11に記載の検出チップ。
- 前記第1枠体は、前記支持体から離れるにつれて前記第1貫通孔の高さ方向に直交する方向における前記第1貫通孔の断面積が大きくなるテーパー部を有する、請求項11または請求項12に記載の検出チップ。
- 前記第2枠体は、前記支持体から離れるにつれて前記第2貫通孔の高さ方向に直交する方向における前記第2貫通孔の断面積が大きくなるテーパー部を有する、請求項11~13のいずれか一項に記載の検出チップ。
- 前記第2貫通孔の高さは、前記第1貫通孔の高さより高い、請求項11~14のいずれか一項に記載の検出チップ。
- 前記第2枠体の前記テーパー部の内壁面は、前記第2貫通孔内に露出した前記支持体の表面の中心から出射され、かつ20°以内の出射角で出射された光線を遮断しないように傾斜している、請求項14に記載の検出チップ。
- 前記第2貫通孔の内壁面は、ブロッキング処理されている、請求項11~16のいずれか一項に記載の検出チップ。
- 前記支持体、前記第1枠体および前記第2枠体のうち少なくとも1つは、試薬を収容するための試薬ホルダーを有する、請求項11~17のいずれか一項に記載の検出チップ。
- 前記第1枠体および前記第2枠体のうち少なくとも1つは、樹脂フィルムからなる、請求項11~18のいずれか一項に記載の検出チップ。
- 前記支持体は、一面上に金属膜が形成された、誘電体からなるプリズムであり、
前記第1枠体および前記第2枠体は、前記一面上に配置され、
前記捕捉体は、前記第1枠体の前記第1貫通孔の内壁面の少なくとも一部と、前記第1貫通孔に露出した前記金属膜の表面とに固定化されている、
請求項11~19のいずれか一項に記載の検出チップ。
Priority Applications (4)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP2017514051A JP6733664B2 (ja) | 2015-04-22 | 2016-04-05 | 検出チップの製造方法および検出チップ |
EP16782988.6A EP3287769A4 (en) | 2015-04-22 | 2016-04-05 | Method of manufacturing sensing chip and sensing chip |
US15/558,506 US10677731B2 (en) | 2015-04-22 | 2016-04-05 | Method of manufacturing sensing chip and sensing chip |
US16/860,808 US20200256796A1 (en) | 2015-04-22 | 2020-04-28 | Method of manufacturing sensing chip and sensing chip |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP2015-087588 | 2015-04-22 | ||
JP2015087588 | 2015-04-22 |
Related Child Applications (2)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US15/558,506 A-371-Of-International US10677731B2 (en) | 2015-04-22 | 2016-04-05 | Method of manufacturing sensing chip and sensing chip |
US16/860,808 Division US20200256796A1 (en) | 2015-04-22 | 2020-04-28 | Method of manufacturing sensing chip and sensing chip |
Publications (1)
Publication Number | Publication Date |
---|---|
WO2016170967A1 true WO2016170967A1 (ja) | 2016-10-27 |
Family
ID=57144597
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/JP2016/061112 WO2016170967A1 (ja) | 2015-04-22 | 2016-04-05 | 検出チップの製造方法および検出チップ |
Country Status (4)
Country | Link |
---|---|
US (2) | US10677731B2 (ja) |
EP (1) | EP3287769A4 (ja) |
JP (1) | JP6733664B2 (ja) |
WO (1) | WO2016170967A1 (ja) |
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPWO2018150943A1 (ja) * | 2017-02-15 | 2019-12-12 | コニカミノルタ株式会社 | 送液システム、検査システム及び送液方法 |
WO2019232858A1 (zh) * | 2018-06-08 | 2019-12-12 | 清华大学 | 具有纳米孔阵的法布里-珀罗结构、制备方法和操作方法 |
Families Citing this family (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2017082196A1 (ja) * | 2015-11-13 | 2017-05-18 | コニカミノルタ株式会社 | 検査チップおよび検査システム |
JP2022072369A (ja) * | 2020-10-29 | 2022-05-17 | 日本碍子株式会社 | 接合体および接合体の製造方法 |
Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPH10239233A (ja) * | 1997-02-26 | 1998-09-11 | Fuji Photo Film Co Ltd | 表面プラズモンセンサー |
JP2000515966A (ja) * | 1996-07-11 | 2000-11-28 | イーツェーベー インスティテュート ファー ヒェモ−ウント ビオゼンゾリック ミュンスター エー.ファー. | 定量的蛍光マーク・アフィニティ・テストを行うための装置および方法 |
WO2005022155A1 (ja) * | 2003-08-29 | 2005-03-10 | Kabushiki Kaisha Toshiba | 発色試薬、濃度測定用キット、濃度測定方法及びそれに用いるセンサチップ |
JP2011503536A (ja) * | 2007-11-05 | 2011-01-27 | コーニンクレッカ フィリップス エレクトロニクス エヌ ヴィ | マイクロエレクトロニクスセンサ |
WO2012157403A1 (ja) * | 2011-05-19 | 2012-11-22 | コニカミノルタホールディングス株式会社 | 表面プラズモン励起増強蛍光測定装置およびこれを用いた蛍光検出方法 |
Family Cites Families (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
DE69830529T2 (de) * | 1997-02-07 | 2006-05-11 | Fuji Photo Film Co., Ltd., Minami-Ashigara | Oberflächen-Plasmonen-Sensor |
US20020127706A1 (en) * | 2001-01-25 | 2002-09-12 | Fuji Photo Film Co., Ltd. | Surface plasmon resonance measuring chip and method of manufacture thereof |
DE60213056T2 (de) | 2001-12-25 | 2007-01-04 | Fuji Photo Film Co., Ltd., Minami-Ashigara | Sensorsystem mit evaneszenten Wellen |
US20040091397A1 (en) * | 2002-11-07 | 2004-05-13 | Corning Incorporated | Multiwell insert device that enables label free detection of cells and other objects |
US8450056B2 (en) * | 2008-05-02 | 2013-05-28 | University Of Rochester | Arrayed imaging reflectometry (AIR) sensor chip comprising virus-like particles suitable for the detection of antiviral immune responses |
JP2013076673A (ja) | 2011-09-30 | 2013-04-25 | Fujifilm Corp | 測定装置 |
JP6026262B2 (ja) * | 2012-12-19 | 2016-11-16 | 富士フイルム株式会社 | 測定装置および測定方法 |
-
2016
- 2016-04-05 JP JP2017514051A patent/JP6733664B2/ja not_active Expired - Fee Related
- 2016-04-05 WO PCT/JP2016/061112 patent/WO2016170967A1/ja active Application Filing
- 2016-04-05 US US15/558,506 patent/US10677731B2/en active Active
- 2016-04-05 EP EP16782988.6A patent/EP3287769A4/en not_active Withdrawn
-
2020
- 2020-04-28 US US16/860,808 patent/US20200256796A1/en not_active Abandoned
Patent Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2000515966A (ja) * | 1996-07-11 | 2000-11-28 | イーツェーベー インスティテュート ファー ヒェモ−ウント ビオゼンゾリック ミュンスター エー.ファー. | 定量的蛍光マーク・アフィニティ・テストを行うための装置および方法 |
JPH10239233A (ja) * | 1997-02-26 | 1998-09-11 | Fuji Photo Film Co Ltd | 表面プラズモンセンサー |
WO2005022155A1 (ja) * | 2003-08-29 | 2005-03-10 | Kabushiki Kaisha Toshiba | 発色試薬、濃度測定用キット、濃度測定方法及びそれに用いるセンサチップ |
JP2011503536A (ja) * | 2007-11-05 | 2011-01-27 | コーニンクレッカ フィリップス エレクトロニクス エヌ ヴィ | マイクロエレクトロニクスセンサ |
WO2012157403A1 (ja) * | 2011-05-19 | 2012-11-22 | コニカミノルタホールディングス株式会社 | 表面プラズモン励起増強蛍光測定装置およびこれを用いた蛍光検出方法 |
Non-Patent Citations (1)
Title |
---|
See also references of EP3287769A4 * |
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPWO2018150943A1 (ja) * | 2017-02-15 | 2019-12-12 | コニカミノルタ株式会社 | 送液システム、検査システム及び送液方法 |
WO2019232858A1 (zh) * | 2018-06-08 | 2019-12-12 | 清华大学 | 具有纳米孔阵的法布里-珀罗结构、制备方法和操作方法 |
Also Published As
Publication number | Publication date |
---|---|
US10677731B2 (en) | 2020-06-09 |
EP3287769A1 (en) | 2018-02-28 |
JP6733664B2 (ja) | 2020-08-05 |
US20180080872A1 (en) | 2018-03-22 |
JPWO2016170967A1 (ja) | 2018-02-15 |
US20200256796A1 (en) | 2020-08-13 |
EP3287769A4 (en) | 2018-04-18 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
US20200256796A1 (en) | Method of manufacturing sensing chip and sensing chip | |
JP6369533B2 (ja) | 測定方法および測定装置 | |
US20200003688A1 (en) | Surface Plasmon Resonance Fluorescence Analysis Device And Surface Plasmon Resonance Fluorescence Analysis Method | |
JP6587024B2 (ja) | 検出方法および検出装置 | |
JP6424890B2 (ja) | 表面プラズモン増強蛍光測定方法、表面プラズモン増強蛍光測定装置および分析チップ | |
JP6856074B2 (ja) | 測定方法、測定装置および測定システム | |
JP2009204483A (ja) | センシング装置 | |
JP6421821B2 (ja) | 検出装置 | |
JP6848975B2 (ja) | 測定方法 | |
JP6760384B2 (ja) | 測定方法 | |
JP6954116B2 (ja) | 測定方法、測定装置および測定チップ | |
JP6717201B2 (ja) | 検出方法および検出装置 | |
JP2009204484A (ja) | センシング装置 | |
WO2016125614A1 (ja) | 検出方法、検出装置およびチップ | |
WO2021009995A1 (ja) | 検出装置および検出方法 | |
WO2015152198A1 (ja) | 表面プラズモン共鳴蛍光分析方法、表面プラズモン共鳴蛍光分析装置および位置合わせ方法 | |
JP6221785B2 (ja) | 検出装置および検出方法 | |
WO2016147774A1 (ja) | 測定方法および測定装置 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
121 | Ep: the epo has been informed by wipo that ep was designated in this application |
Ref document number: 16782988 Country of ref document: EP Kind code of ref document: A1 |
|
ENP | Entry into the national phase |
Ref document number: 2017514051 Country of ref document: JP Kind code of ref document: A |
|
WWE | Wipo information: entry into national phase |
Ref document number: 15558506 Country of ref document: US |
|
REEP | Request for entry into the european phase |
Ref document number: 2016782988 Country of ref document: EP |
|
NENP | Non-entry into the national phase |
Ref country code: DE |