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WO2013039065A1 - Capteur de fluorescence et procédé de mesure de composant d'analyte - Google Patents

Capteur de fluorescence et procédé de mesure de composant d'analyte Download PDF

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Publication number
WO2013039065A1
WO2013039065A1 PCT/JP2012/073195 JP2012073195W WO2013039065A1 WO 2013039065 A1 WO2013039065 A1 WO 2013039065A1 JP 2012073195 W JP2012073195 W JP 2012073195W WO 2013039065 A1 WO2013039065 A1 WO 2013039065A1
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Prior art keywords
light
fluorescent
indicator layer
fluorescence
photodiode
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English (en)
Japanese (ja)
Inventor
松本　淳
悦朗 清水
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Terumo Corp
Olympus Corp
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Terumo Corp
Olympus Corp
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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
    • G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"

Definitions

  • the present invention relates to a fluorescent sensor and an analyte component measuring method for measuring an analyte component in a living body.
  • various analyzers have been developed as devices for quantifying the concentration of an analyte in a liquid.
  • one of such analyzers injects several milliliters of an analyte-containing solution into a transparent container of a certain volume, irradiates the container with appropriate light, and measures the light absorption and fluorescence of the analyte. Is done.
  • Such an analyzer is called a fluorescence spectrophotometer.
  • Such a fluorescent sensor has been proposed that has been miniaturized by recent semiconductor device manufacturing technology and micromachining technology.
  • a photodiode is formed on a thin transparent substrate, an indicator layer that reacts with an analyte to emit fluorescence is disposed on the photodiode, and on both sides and an upper portion of the photodiode.
  • a fluorescent sensor configured to guide excitation light to an indicator layer on a photodiode by providing an optical path.
  • the excitation light is irradiated from the back surface direction of the photodiode to the entire indicator layer so that the excitation light is not directly irradiated toward the light receiving surface of the photodiode.
  • the fluorescent compound in the indicator layer deteriorates when exposed to excitation light and does not emit fluorescence. For this reason, a fluorescence sensor that can keep the lifetime of the fluorescence sensor longer is desired.
  • An object of the present invention is to provide a fluorescent sensor capable of extending the lifetime more than ever.
  • the fluorescent sensor of the present invention detects an analyte component of a living body, and has an indicator layer fixed with a fluorescent substance that emits fluorescence according to the amount of the analyte component when irradiated with excitation light;
  • a photodiode that receives the fluorescence and outputs an electrical signal corresponding to the intensity of the fluorescence, and a light source that irradiates the indicator layer with the excitation light, the indicator layer on the photodiode The light source irradiates the excitation light into the indicator layer in a direction parallel to the light receiving surface of the photodiode.
  • the analyte component measuring method of the present invention for achieving the above object is characterized in that the above-mentioned light source is provided in the indicator layer in which a fluorescent substance that emits fluorescence according to the amount of the analyte component in the living body is fixed by the excitation light.
  • An analyte component measuring method in which excitation light is incident, fluorescence emitted from the fluorescent material is received and the intensity of the fluorescence is measured, wherein a site where the fluorescent material generates fluorescence is away from the light source And the intensity of fluorescence emitted from the moved site is measured.
  • the excitation light is irradiated toward the indicator layer in parallel with the photodiode light receiving surface.
  • the fluorescent material fixed in the indicator layer gradually becomes transparent from the light source to the back, and the position that emits the fluorescence moves (shifts) most. For this reason, it becomes possible to continue the measurement until the excitation light reaches the innermost part from the light source of the indicator layer, and the life of the indicator layer can be extended.
  • the excitation light is incident on the fluorescent material fixed in the indicator layer so that the position where the fluorescence is emitted is farthest from the light source. It was decided to move (shift). For this reason, the measurement can be continued until the excitation light reaches the innermost part from the light source of the indicator layer, and the lifetime of the indicator layer can be extended.
  • (A) is sectional drawing
  • (B) is a top view.
  • 3 is a graph showing transmittance characteristics of a polysilicon film and a silicon carbide film.
  • It is a fragmentary sectional view of the light source part for demonstrating the case where LED is used for a light source.
  • It is an external appearance perspective view of a sensor system.
  • FIG. 1 is a schematic diagram of the fluorescent sensor of the present embodiment
  • FIG. 1A is a schematic cross-sectional view of the fluorescent sensor
  • FIG. 1B is a schematic diagram of FIG. 1A excluding the light shielding film of the fluorescent sensor. It is a schematic plan view seen from above.
  • the fluorescent sensor 1 includes a photodiode 3 provided on the surface of the substrate 2, an optical filter 4 that covers at least the light receiving surface of the photodiode 3, and an indicator layer that is located on the photodiode 3 and provided on the optical filter 4. 5, a light source 6 that emits excitation light to the indicator layer 5 in a direction parallel to the light receiving surface on the side of the photodiode 3, and a light shielding film 7 that covers at least the indicator layer 5.
  • the substrate 2 is, for example, a silicon single crystal substrate.
  • a photodiode 3 is directly formed on the surface of the substrate 2 by a planar technique which is one of semiconductor device manufacturing techniques. Referring to FIG. 1B, one wiring 51 for detecting a signal from the photodiode 3 extends to the substrate end on the light source 6 side on the substrate surface. The other signal line of the photodiode 3 can be taken from the substrate 2. Further, power supply wirings 52 and 53 to the light source 6 are also formed on the substrate surface. In addition, arrangement
  • the optical filter 4 has a property of blocking only the light source wavelength so that the light from the light source 6 does not directly enter the photodiode, while transmitting the fluorescent wavelength from the indicator layer 5.
  • the arrangement position may be any position that covers at least the photodiode 3, but it is preferable that the arrangement position is just below the light source in consideration of irregular reflection of light inside the apparatus. This is to prevent the light emitted from the light source 6 from entering directly into the substrate 2 and affecting the photodiode 3.
  • an optical filter 4 for example, a light source 6 having an excitation light wavelength of 350 nm to 420 nm, preferably a central wavelength of 375 nm is used, while the indicator layer 5 has a fluorescence wavelength of 400 nm to 600 nm and a peak wavelength of 480 nm.
  • the wavelength of 350 nm to 420 nm may be blocked, while 420 nm or more, preferably 450 nm to 600 nm may be transmitted.
  • a filter comprising a silicon oxide film having a thickness of several tens to several hundreds nm and a polysilicon film having a thickness of several hundreds nm to several ⁇ m formed thereon on the surface of the photodiode element.
  • a layer may be provided.
  • the filter layer configured as described above can suppress the excitation light having an excitation wavelength of 375 nm from entering the photodiode. For example, if a polysilicon thickness of 900 nm, can be suppressed to about 1/10 8. When an LED is used for the light source 6 to be described later, there is slight light emission at other wavelengths of the excitation wavelength.
  • tantalum oxide and silicon oxide having a thickness of several nm to several tens of nm are alternately formed per layer.
  • SiC silicon carbide
  • FIG. 2 is a graph showing transmittance characteristics of the polysilicon film and the silicon carbide film, where the horizontal axis indicates the wavelength of light and the vertical axis indicates the light transmittance.
  • A shows the case of a polysilicon film having a film thickness of 0.5 ⁇ m
  • B shows the case of a silicon carbide film having a film thickness of 360 ⁇ m.
  • both (A) the polysilicon film and (B) the silicon carbide film have a transmittance of 10 ⁇ 7 or less at the wavelength of the excitation light E shorter than 375 nm, whereas at the wavelength of the fluorescence F at 460 nm,
  • the transmittance is 10 -1 or more, that is, 10% or more, and the transmittance selectivity by wavelength is 6 digits or more.
  • the optical filter 4 when a silicon film is used as the optical filter 4, a thickness of 1 ⁇ m is sufficient, so that it can be integrally formed on the semiconductor substrate after the formation of the photodiode 3 in a well-known semiconductor manufacturing process.
  • the silicon of the material of the optical filter 4 may be non-doped, but is preferably a polycrystalline silicon film or an amorphous silicon film having a thickness of sub ⁇ m to several ⁇ m doped with an impurity such as phosphorus.
  • gallium phosphide can also be preferably used as the optical filter 4 because it has a low transmittance at a wavelength of excitation light shorter than around 375 nm and a high transmittance at a wavelength of fluorescence of 460 nm.
  • the indicator layer 5 is provided so as to cover the photodiode 3, and the excitation light from the light source 6 enters the indicator layer in parallel to the light receiving surface of the photodiode 3. Is arranged.
  • the indicator layer 5 preferably covers an area equal to or larger than the area of the photodiode light receiving surface. This is to make it possible to detect as much fluorescence emitted from the indicator layer 5 as possible over the entire detection region of the photodiode 3. The specific size of the indicator layer 5 will be described later.
  • the indicator layer 5 includes a fluorescent substance that emits fluorescence according to the amount of the analyte when it is exposed to excitation light, and a poly (meth) acrylamide residue for fixing and holding the fluorescent substance so as not to move in the indicator layer.
  • a hydrogel or polymer comprising a gel containing a polymerizable monomer is used.
  • the gel or the polymer preferably includes, for example, a fluorescent monomer (fluorescent substance) having a phenylboronic acid at a sugar recognition site, and a fluorescent monomer having a phenylboronic acid group of the following chemical formula 1 It is further preferable to provide a copolymer with a polymerizable monomer containing a (meth) acrylamide residue.
  • X 1 and X 2 may be the same or different, and —COO—, —OCO—, —CH 2 NR—, —NR—, —NRCO—, —CONR—, —SO 2 NR—, -NRSO 2 -, - O -, - S -, - SS -, - NRCOO -, - OCONR- and -CO- is selected from the group consisting of 1 to 30 carbon atoms containing at least one substituent alkylene R is hydrogen or an optionally substituted alkyl group.
  • an alkylene containing at least one kind of substituent refers to an alkylene having a substituent bonded to an end of the alkylene and an alkylene having a substituent in the chain.
  • the number of carbon atoms of the alkylene is preferably 1-30, more preferably 3-12. Specific examples include propylene, hexylene and octylene.
  • the substituent contained in the alkylene is preferably —NRCO— or —CONR—.
  • R is an alkyl group, those having 1 to 10 carbon atoms are preferred, and 1 to 5 is more preferred. Specific examples include a methyl group, an ethyl group, a propyl group, a butyl group, and a pentyl group.
  • R is preferably hydrogen.
  • Z 1 and Z 2 may be the same or different and each represents —O— or —NR′—, and R ′ represents hydrogen or an optionally substituted alkyl group.
  • the alkyl group preferably has 1 to 10 carbon atoms, more preferably 1 to 5 carbon atoms. Specific examples include a methyl group, an ethyl group, a propyl group, a butyl group, and a pentyl group.
  • Z 1 and Z 2 are preferably —N—.
  • Y 1 and Y 2 may be the same or different and are divalent organic residues which may be substituted.
  • Y 1 and Y 2 are preferably hydrophilic enough to make the fluorescent monomer water-soluble.
  • the hydrophilicity to such an extent that the fluorescent monomer can be made water-soluble means that it dissolves in water in the concentration range necessary for polymerizing the fluorescent monomer without the presence of an organic solvent or a solubilizing agent.
  • Y 1 and Y 2 having a hydrophilic group such as amino group, carboxyl group, sulfo group, nitro group, amino group, phosphoric acid group and hydroxyl group, and hydrophilic such as ether bond, amide bond and ester bond in the structure What has a sexual bond can be illustrated.
  • Y 1 and Y 2 preferably include a structure represented by the following chemical formula 2 or chemical formula 3 in the organic residue, and may further have another substituent or a divalent organic residue.
  • n is preferably 2 to 4, more preferably 2 or 3
  • j is preferably 1 to 3, more preferably 1
  • m is preferably 20 to 150, more preferably 40. ⁇ 120.
  • the molecular weight of Y is preferably 500 to 10,000, more preferably 1,000 to 5,000.
  • the divalent organic residue represented by Chemical Formula 2 or Chemical Formula 3 can be prepared, for example, by polymerizing alkylene glycol such as ethylene glycol or propylene glycol, vinyl alcohol, or the like.
  • the fluorescent monomer used in the present embodiment can obtain the following effects more specifically by introducing the hydrophilic chain Y.
  • the fluorescent monomer becomes water-soluble, it is possible to efficiently perform immobilization and polymerization reaction when forming the fluorescent sensor material.
  • an acrylamide gel is prepared, polymerization can be performed using only water as a solvent, and a product having high physical strength, stability, and uniformity can be obtained.
  • an organic solvent or the like which may result in an unfavorable gel.
  • hydrophilic chain changes the environment and motility around phenylboronic acid that interacts with the substance to be detected, contributing to improved sensitivity, accuracy, response speed, and selectivity of the saccharide as the substance to be measured. .
  • the hydrophilic chain stabilizes the entire fluorescent sensor material, for example a polymerized structure. (4) Since it can react only with water, it can superpose
  • L represents an optionally substituted alkyl group having 1 to 10 carbon atoms. Specific examples include a methyl group, an ethyl group, a propyl group, a butyl group, and a pentyl group. More preferably, the alkyl group has 1 to 4 carbon atoms. L is preferably a methyl group.
  • the fluorescent monomer used in the present embodiment is characterized in that Y is introduced into the monomer compound for saccharide detection via the above X, and thereby the physical properties and stability of the fluorescent monomer compound, detection sensitivity, and detection. It is possible to improve the accuracy, the selectivity of the saccharide that is the substance to be measured, and the like.
  • the fluorescent monomer used in this embodiment is a phenylboronic acid derivative containing an anthracene skeleton, and the anthracene skeleton is known to act as a fluorescent atomic group.
  • a phenylboronic acid moiety and a saccharide form a stable complex, it fluoresces due to the presence of a fluorescent group, but the fluorescent monomer used in this embodiment has two phenylboronic acids, Excellent detection sensitivity.
  • the obtained derivative has an acryloyl group and an amide in its structure, and includes (meth) acrylamide and derivatives thereof.
  • acrylamide, methacrylamide, N, N-dimethylacrylamide, N-isopropylacrylamide, N-tert-butylacrylamide, N-tris-hydroxymethylacrylamide, N-hydroxymethylacrylamide, N- (n-butoxymethyl) acrylamide examples include condensates of (meth) acryloyl chlorides such as N-acryloyl lysine and N-acryloyl hexamethylene diamine with compounds having amino acids or active amino groups, and compounds represented by Formula 4.
  • A is hydrogen or a methyl group
  • U and U ′ may be the same or different, and are hydrogen or an optionally substituted alkyl group.
  • alkyl group include a methyl group, an ethyl group, a propyl group, a butyl group, and a pentyl group.
  • a polymer composed of a polymerizable monomer containing a (meth) (meth) acrylamide residue has high hydrophilicity
  • a highly hydrophobic fluorescent group containing phenylboronic acid present in the fluorescent monomer Is incorporated into a highly hydrophilic structure.
  • water-soluble saccharides can easily approach and bind to the fluorescent atomic group.
  • a fluorescent sensor material is used for the indicator layer 5, and the composition of the gel and polymer, the manufacturing method thereof, and the like, which are forms of the fluorescent sensor material, will be described below.
  • the copolymer composition molar ratio ((I) :( II)) of the fluorescent monomer (I) and the polymerizable monomer (II) containing the (meth) acrylamide residue constituting the fluorescent sensor substance is 1: It is preferably 50 to 1: 6,000, more preferably 1: 150 to 1: 3,000.
  • the ratio of the fluorescent monomer is larger than the molar ratio 1:50, the degree of freedom is lost due to the bulk of the fluorescent monomer, and the interaction with the saccharide may be reduced.
  • the ratio of the fluorescent monomer compound is smaller than the molar ratio 1: 6,000, the absolute amount of fluorescence intensity may not be ensured.
  • the weight average molecular weight of the two-component polymer fluorescent sensor substance is preferably 50,000 to 750,000, more preferably 150,000 to 450,000 in terms of polyethylene oxide by GPC.
  • the fluorescent sensor substance used in the present embodiment may use other components in combination.
  • Such components include crosslinkable monomers, other crosslinkable components, cationic monomers that can be cations in water, anionic monomers that can be anions in water, and nonionic monomers that do not have ions.
  • the crosslinkable monomer widely includes those capable of introducing a three-dimensional cross-linked structure into a fluorescent sensor substance by a polymerizable double bond, and varies depending on the substituent of the fluorescent sensor substance to be used, but N, N′-methylenebis ( (Meth) acrylamide, N, N '-(1,2-dihydroxyethylene) -bis (meth) acrylamide, diethylene glycol di (meth) acrylate, (poly) ethylene glycol di (meth) acrylate, triethylene glycol di (meth) acrylate , Propylene glycol di (meth) acrylate, trimethylolpropane di (meth) acrylate, trimethylolpropane tri (meth) acrylate, pentaerythritol di (meth) acrylate, pentaerythritol tri (meth) acrylate, pentaerythritol Tetra (meth) acrylate, (poly) propylene glycol di (
  • crosslinkable components widely include compounds having two or more functional groups, and depending on the substituent of the fluorescent sensor material used, triallyl cyanurate, triallyl isocyanurate, triallyl phosphate, triallylamine , Poly (meth) allyloxyalkane, (poly) ethylene glycol diglycidyl ether, glycerol diglycidyl ether, ethylene glycol, polyethylene glycol, propylene glycol, glycerin, pentaerythritol, ethylenediamine, polyethyleneimine, glycidyl (meth) acrylate, Mention may also be made of triallyl isocyanurate, trimethylolpropane di (meth) allyl ether, tetraallyloxyethane or glycerol propoxytriacrylate. Two or more of these may be used in combination.
  • Examples of the cationic monomer that can become a cation in water include dimethylaminoethyl (meth) acrylate, diethylaminoethyl (meth) acrylate, and 4-vinylpyridine. Two or more of these may be used in combination.
  • Nonionic monomers having no ions include 2-hydroxyethyl (meth) acrylate, 3-methoxypropyl (meth) acrylate, 4-hydroxybutyl (meth) acrylate, 2-methoxyethyl acrylate or 1,4-cyclohexanedimethanol A monoacrylate etc. can be mentioned. Two or more of these may be used in combination.
  • crosslinkable monomers other crosslinkable components, cationic monomers, anionic monomers and nonionic monomers may be used in combination of two or more.
  • the blending amount of these other components is preferably from 0.1 to 10 mol%, more preferably from 2 to 7 mol% of the total amount of the fluorescent monomer compound and the polymerizable monomer containing a (meth) acrylamide residue. .
  • a three-dimensional crosslinked structure can be formed, and hydrophilicity adjustment, introduction of a reaction starting point, and the like can be performed. The three-dimensional crosslinked structure will be described later.
  • the fluorescent sensor substance (fluorescent substance) used in the present embodiment preferably has a structure represented by Chemical Formula 5.
  • X 1 , X 2 , Z 1 , Z 2 , Y 1 , Y 2 , and L are the same as the fluorescent monomer shown in Chemical Formula 1.
  • U 1, U 2, U 3 , U 4, A 1 and A 2 are the same as the polymerizable monomer containing a shown in Chemical Formula 4 (meth) acrylamide residual group.
  • the molar ratio of p 1 and q 1 (p 1 : q 1 ) and the molar ratio of p 2 and q 2 (p 2 : q 2 ) correspond to the molar ratio of (I) :( II).
  • the ratio is preferably 1:50 to 1: 6,000, more preferably 1: 150 to 1: 3,000.
  • the fluorescent sensor substance (fluorescent substance) used in the present embodiment may have a three-dimensional cross-linked structure in which at least a part of the copolymer forms an intermolecular cross-link.
  • the fluorescent monomer When a three-dimensional crosslink is formed on the poly (meth) acrylamide chain, the fluorescent monomer is fixed to the substrate, and sugars can be easily detected without eluting the fluorescent monomer compound even in an aqueous solution.
  • the fluorescent monomer has a hydrophobic moiety that emits fluorescence by binding to a saccharide, and the hydrophobic moiety is bonded to a poly (meth) acrylamide chain via a divalent organic residue represented by Y. Therefore, the freedom degree which can be couple
  • fluorescent monomer compound As the fluorescent monomer represented by Chemical Formula 1, a compound in which X is —C 6 H 12 —NHCO—, Y is a PEG residue, Z is —O—, and L is —CH 3 group, An example of a method for producing 9,10-bis (methylene) [[N- (orthoboronobenzyl) methylene] -N-[(acryloylpolyoxyethylene) carbonylamino] -n-hexamethylene] -2-acetylanthracene This will be described below.
  • 2-acetyl-9,10-dimethylanthracene as a raw material and heating the carbon tetrachloride / chloroform solvent to react with N-bromosuccinimide (NBS) and benzoyl peroxide (BPO), 2-acetyl- It is 9,10-bis (bromomethylene) anthracene.
  • NBS N-bromosuccinimide
  • BPO benzoyl peroxide
  • the target product can be obtained by reacting acryloyl- (polyethylene glycol) -N-hydroxysuccinimide ester in a basic buffer.
  • Z is NH
  • acrylamide- (polyethylene glycol) -N-hydroxysuccinimide ester may be used instead of acryloyl- (polyethylene glycol) -N-hydroxysuccinimide ester used in the last step.
  • the blending amount is preferably 0.1 to 10 mol%, more preferably 2 to 7 mol% of the total amount of the fluorescent monomer compound and the polymerizable monomer containing a (meth) acrylamide residue. %.
  • it is preferably added at the same time as the polymerization initiator and the polymerization accelerator during the polymerization.
  • polymerization initiator examples include persulfates such as ammonium persulfate, sodium persulfate, and potassium persulfate; hydrogen peroxide; azo compounds such as azobis-2-methylpropionamidine hydrochloride or azoisobutyronitrile; benzoyl Peroxides such as peroxide, lauroyl peroxide, cumene hydroperoxide or benzoyl oxide can be used, and one or more of these can be used.
  • persulfates such as ammonium persulfate, sodium persulfate, and potassium persulfate
  • hydrogen peroxide such as azobis-2-methylpropionamidine hydrochloride or azoisobutyronitrile
  • benzoyl Peroxides such as peroxide, lauroyl peroxide, cumene hydroperoxide or benzoyl oxide can be used, and one or more of these can be used.
  • a reducing agent such as sodium hydrogen sulfite, sodium sulfite, molle salt, sodium pyrobisulfite, formaldehyde sodium sulfoxylate or ascorbic acid; ethylenediamine, ethylenediaminesodium tetraacetate, glycine or N, N, N ′ , N′-tetramethylethylenediamine and other amine compounds; and the like can be used in combination.
  • the polymerization temperature is preferably 15 to 75 ° C., more preferably 20 to 60 ° C., and the polymerization time is 1 to 20 hours, more preferably 2 to 8 hours.
  • the compound represented by Chemical Formula 5 can also be produced without copolymerization of the fluorescent monomer represented by Chemical Formula 1 and a polymerizable monomer containing a (meth) acrylamide residue. Since the fluorescent monomer represented by the chemical formula 1 is synthesized in a plurality of steps, even if another compound is allowed to act on the intermediate product without using the fluorescent monomer represented by the compound 1 as a raw material, the fluorescent sensor finally represented by the chemical formula 5 A substance can be produced.
  • a fluorescent sensor substance represented by Chemical Formula 5 can also be produced by allowing a compound introduced with a carboxyl group to act in the presence of a coupling reagent.
  • a fluorescent sensor represented by Chemical Formula 5 can be obtained by polymerizing a polymerizable monomer having a (meth) acrylamide residue in advance and then copolymerizing with the above fluorescent monomer compound in the presence of a polymerization initiator or a polymerization accelerator. A substance can be produced.
  • the fluorescent sensor material used in the present embodiment may have three-dimensional cross-linking, but the method for introducing the three-dimensional cross-linking is not limited. There is a method in which a cross-linking component is allowed to act on the fluorescent sensor material used in the present embodiment to form intermolecular cross-links on at least a part of the fluorescent sensor material and the fluorescent sensor material.
  • a three-dimensional crosslinking can be formed even if a crosslinking component is added to the reaction solvent. it can.
  • a fluorescent sensor substance when used as a saccharide measuring sensor for implantation in the body, it is generally fixed to a base material in order to prevent the fluorescent sensor substance from flowing out.
  • a polymerizable monomer containing a (meth) acrylamide residue or a polymer thereof a fluorescent monomer compound represented by Chemical Formula 1 is optionally used with a crosslinking component, and these are polymerized to be fixed to a substrate. Three-dimensional crosslinking can be performed simultaneously.
  • cross-linkable monomer As such a cross-linking component, the cross-linkable monomer, other cross-linkable component, cationic monomer, anionic monomer and nonionic monomer described in the section of other components which can be blended with the fluorescent sensor substance are preferably used.
  • the crosslinkable monomer and other crosslinkable components can be used more preferably. Two or more of these may be used in combination.
  • Such an optical characteristic of the indicator layer 5 absorbs the light of the excitation light and is basically transparent to the light having the fluorescence wavelength.
  • the fluorescent substance in the indicator layer 5 deteriorates when it is irradiated with excitation light, and the ability to absorb the excitation light also decreases at the same time as the fluorescence energy gradually decreases. That is, the fluorescent substance contained in the indicator layer 5 has a characteristic that when the excitation light hits, the fluorescence emission ability decreases with the passage of time while the light transmission increases.
  • the portion close to the light source 6 in the indicator layer 5 becomes easy to transmit excitation light (that is, it becomes difficult to absorb excitation light) and does not emit fluorescence. Come.
  • the excitation light reaches a position distant from the light source 6 because the part that is deteriorated and transmits increases. If it does so, the part which absorbs light and emits fluorescence will move.
  • the lifetime of the fluorescence sensor can be kept long by using such characteristics.
  • an adhesive layer made of a silane coupling agent (not shown) is applied on the optical filter 4, and a hydrogel is bonded thereto.
  • a glass layer is laminated on the optical filter 4 by CVD to block water from entering, and then an alcohol aqueous solution of 0.2% aminopropyltriethoxysilane (silane coupling agent). To form an amino group. Further, after contacting with 0.1% acryloyl chloride methylene chloride solution to form acrylamide group, fluorescent monomer (concentration of about 1% to 10%) having acrylamide terminal on this surface and acrylamide monomer (5% to 20%) To a final concentration of 0.4% sodium persulfate and 0.04% tetraethylmethylenediamine.
  • the concentration of sodium persulfate and tetraethylmethylenediamine may be appropriately changed in order to change the polymerization rate during the production process.
  • the indicator layer 5 fluorescent gel
  • the method for forming the indicator layer 5 is not limited to the above-exemplified method, and various other methods may be used.
  • the light source 6 is, for example, an LED. As shown in FIG. 1A, the excitation light from the light source 6 enters the indicator layer 5 in a direction parallel to the light receiving surface of the photodiode (that is, in FIG. 1A, Excitation light is incident from the light source 6 to the indicator layer 5 from the side (in the direction of arrow A shown).
  • FIG. 3 is a cross-sectional view of a light source portion for explaining a case where an LED is used as a light source, where (A) shows a first example and (B) shows a second example.
  • an LED 61 is manufactured on a substrate 2 so that light is emitted in a vertical direction with respect to the substrate surface, and a prism 62 is provided on the LED 61. It is arranged.
  • An angle filter 63 is provided on the light emission side end face of the prism 62.
  • a wiring layer 64 is formed.
  • the other electrode of the LED 61 is connected to another wiring provided on the substrate 2 by a bonding wire (not shown).
  • the LED 61 provided on the substrate 2 emits light in both the prism 62 direction and the substrate 2 direction.
  • the light emitted toward the prism 62 is emitted toward the indicator layer 5 by the prism 62.
  • the light emitted toward the substrate 2 is reflected by the metal of the wiring layer 64 itself, the light is emitted toward the prism 62 after all. Therefore, most of the light emitted from the LED 61 is emitted in the direction of the indicator layer 5 (in the direction of the arrow in the figure) through the prism 62.
  • a reflective film may be further sandwiched between the wiring layer 64 and the LED 61.
  • the prism 62 is made of glass, transparent resin, or the like.
  • the angle filter 63 is a filter that selects and transmits only light incident in a predetermined angle range.
  • the light-shielding material is processed into a louver structure by microfabrication and coated with a transparent resin.
  • the light shielding material is preferably a metal or a semiconductor material having a low transmittance.
  • a light transmissive material such as a resin may be coated with a light shielding film such as a metal thin film, or carbon black may be included.
  • the light shielding may be realized by the means.
  • a lens array such as a SELFOC lens or a fiber array structure
  • the fiber array can be realized as a structure in which clad-core type optical fiber structures are arranged in an array or a structure in which a light-transmitting thin tube array is covered with a reflector.
  • a pedestal 65 that supports the back surface of the LED is formed in advance on the LED mounting portion on the substrate 2, and the light emission direction of the LED 61 is on the surface 66 side.
  • the LED 61 is arranged so as to face the indicator layer 5.
  • the surface 66 is provided with a wiring layer 67 connected to one electrode of the LED 61 (actually one conductive layer (P layer or N layer) of the LED chip) so as to cover the entire surface.
  • the other electrode of the LED 61 is connected to another electrode wiring provided on the substrate 2 by a bonding wire (not shown).
  • one of the light emission surfaces of the LED 61 is in the direction of the indicator layer 5, and the other is in the opposite direction from the indicator layer 5.
  • the light emitted in the opposite direction is reflected by the wiring layer 67 (metal) itself provided on the surface 66 and is eventually emitted to the indicator layer 5. Therefore, most of the light emitted from the LED 61 is emitted in the indicator layer direction (arrow direction in the figure).
  • a reflective film may be provided on the surface 66 in addition to the wiring layer 67.
  • the light source 6 in addition to the LED 61, for example, a semiconductor laser chip can be used. In that case, the structure shown in FIG.
  • the excitation light may be guided from the separately provided light source 6 by an optical fiber.
  • FIG. 4 is a partial cross-sectional view showing an example in which an optical fiber is placed as the light source 6 of excitation light on the substrate 2.
  • the emission end of the optical fiber 68 faces the indicator layer 5 and the light emitted from the optical fiber 68 faces the indicator layer 5 as shown in the figure. (Arrow direction shown).
  • a collimator lens (not shown) is preferably disposed at the exit end of the optical fiber 68. Thereby, most of the light emitted from the optical fiber 68 can be emitted into the indicator layer 5 as parallel light. Even in this case, it is preferable to provide the optical filter 4 so that irregular reflection in the apparatus does not directly enter the photodiode.
  • the light shielding film 7 shields light entering the fluorescent sensor 1 from the outside of the fluorescent sensor 1 and transmits the substance to be measured.
  • an optical separation layer disclosed in JP-A-2005-315871 can be used. This optical separation layer comprises an opaque substance and an optical separation layer substrate carrying the opaque material.
  • Opaque materials do not transmit and reflect light such as ultraviolet light, visible light, and infrared light. Also, it has a function of not emitting excitation light outside the sensor.
  • an opaque substance for example, carbon black, fullerene, carbon nanotube, or the like can be used.
  • the substrate for the optical separation layer various polymer substances can be used, but the polymer substance may be crosslinked or modified.
  • the hydrophilic polymer substance for example, dextran, polyacrylamide, polyethylene glycol, polyvinyl alcohol, polyhydroxyethyl methacrylate, or a copolymer thereof can be used.
  • a polymer porous membrane formed by a phase inversion method can be used as the substrate for the optical separation layer.
  • the phase inversion method for obtaining a porous polymer membrane for an optical separation layer substrate is a conventionally known method, and for example, a wet film forming method can be used. Examples of this material include methyl cellulose, polyether sulfone, polysulfone, polyethylene, and polyurethane.
  • the average pore diameter of such a polymer porous membrane is preferably 0.001 ⁇ m to 0.1 ⁇ m. More preferably, the thickness is 0.005 ⁇ m to 0.01 ⁇ m. When it is desired to accelerate the response speed, the average pore diameter is more preferably set to 0.01 ⁇ m to 0.05 ⁇ m, for example.
  • the photodiode 3 and the wiring layer are formed on the main surface side of the silicon substrate.
  • the wiring layer is a wiring for taking out a signal from the photodiode 3 to the outside and a power supply wiring for supplying power to the LED 61.
  • the wiring here is made of aluminum, polysilicon, or the like.
  • the optical filter 4 described above is formed on the photodiode 3.
  • an insulating film for example, SiO 2
  • a contact hole for connecting to the power supply wiring is formed in the LED forming portion on the insulating film, and the power supply wiring and the LED 61 are connected.
  • various methods such as flip chip, solder, a metal-metal direct bonding method, and a method using a conductive resin can be used.
  • the indicator layer 5 and the light shielding film 7 are formed. Then, a resin film is formed so as to cover the whole, and the resin film is opened only at the portion of the light shielding film 7 to expose the surface of the light shielding film 7. Similarly, the pad portion is also exposed. As a result, a chip-shaped fluorescent sensor is completed.
  • a photodiode, a wiring layer, and an optical filter are sequentially formed using a flexible insulating substrate such as a resin. Specifically, it is as follows.
  • a silicon layer is formed on a flexible insulating substrate such as a resin.
  • the silicon layer is preferably single crystal silicon in order to form a highly sensitive photodiode, but may be polysilicon or amorphous silicon as long as sufficient sensitivity can be obtained as a fluorescent sensor.
  • These silicon layers may be formed directly on an insulating substrate by CVD or the like, or a silicon single crystal substrate may be attached.
  • the photodiode 3 is formed on the formed silicon layer. Thereafter, the portion of the insulating substrate where the photodiode 3 is not present (the portion where the silicon layer is not present) is planarized using a resin material such as polyimide. Then, a thin film of aluminum or gold for forming a wiring layer is formed on the photodiode 3 and the planarizing film by vapor deposition or sputtering, and the wiring of the photodiode is patterned by a photolithography process and etching. At this time, as a matter of course, unnecessary films on the photodiode light-receiving surface are removed.
  • an insulating thin film is coated on the wiring by sputtering, vapor deposition, CVD (Chemical Vapor deposition), etc., and the wiring is insulated.
  • this coating film is formed with a uniform thickness, a concave portion is formed between the wirings.
  • the wirings can be insulated. At the same time, the unevenness is filled and a smooth (flat) surface is obtained.
  • An insulating resin may be coated instead of the metal alkoxide.
  • the space for the indicator layer 5 is put in a secured mold material, the resin is injected and cured, and the indicator material is When introduced and sealed with a light-shielding film 7, the fluorescent sensor 1 is completed.
  • a photodiode is separately formed using a silicon substrate and this is attached to the insulating substrate. Specifically, it is as follows.
  • a wiring layer for the photodiode is formed on the insulating substrate.
  • the formed wiring layer is covered and flattened with polyimide, metal alkoxide, etc. in order to eliminate unevenness due to wiring, and bumps are formed only on the portion connected to the photodiode, and then the photodiode is electrically connected. Make sure you can connect.
  • the photodiode 3 is formed separately using a silicon substrate (preferably a single crystal silicon substrate).
  • a silicon substrate preferably a single crystal silicon substrate.
  • a plurality of photodiodes are formed on a single silicon substrate in the same manner as in a normal semiconductor element manufacturing method.
  • the optical filter 4 is formed on the entire surface of the formed photodiode.
  • the optical filter 4 may be formed using the various materials already described.
  • a pad or the like is formed on the photodiode formed on the silicon substrate so that electrical connection can be obtained from the surface opposite to the optical filter forming surface.
  • the photodiode is cut out from the silicon substrate into a chip (for example, flip chip). As a result, a photodiode chip with an optical filter is completed. If necessary, the silicon substrate (or chip) after forming the photodiode may be polished and thinned.
  • the silicon substrate on which the photodiode is formed may be polished and thinned to form the optical filter on the back side.
  • the photodiode formation surface of the silicon substrate is the front surface
  • the back surface side of the thinned silicon substrate becomes the light receiving surface of the photodiode, and an optical filter is formed on this light receiving surface.
  • pads for electrical wiring are formed on the surface side when the photodiode is formed).
  • the photodiode chip with an optical filter is attached while being connected to the wiring layer of the insulating substrate (the photodiode chip pad and the wiring bump are connected).
  • the space for the indicator layer 5 is put in a secured mold, the resin is injected and cured, the indicator material is introduced, and the light shielding film 7 If sealed, the fluorescent sensor 1 is completed.
  • the wiring layer may be formed later after the photodiode chip with an optical filter is attached to the insulating substrate.
  • the optical filter may be formed with a contact hole for electrical connection with the photodiode.
  • a photodiode may be formed by providing a silicon layer thereon.
  • the photodiode may be an organic semiconductor as well as an inorganic semiconductor.
  • it can be manufactured using various technologies used in semiconductor element manufacturing processes and MEMS (Micro Electro Mechanical System).
  • FIG. 5 is an explanatory diagram for explaining the operation of the fluorescence sensor 1.
  • FIG. 5A shows the initial stage of measurement.
  • the larger the black arrow the stronger the excitation light (the same applies to FIG. 5B).
  • the larger the white arrow the more fluorescence is emitted (the same applies to FIG. 5B).
  • the excitation light is almost absorbed on the side closer to the light source, and the excitation light does not reach the farther from the light source 6. Almost no excitation light reaches the part farthest from the light source 6. In this initial stage, the fluorescence increases near the light source 6.
  • the excitation light reaches far from the light source 6. In the figure, it reaches strongly near the center of the indicator layer 5 and only weakly reaches farther away. The position where fluorescence is emitted most is far from the light source 6 and increases near the center. In addition, although excitation light passes strongly at a position close to the light source 6, almost no fluorescence is emitted.
  • the analyte can be measured over a long period of time.
  • excitation light is incident from a light source into an indicator layer to which a fluorescent material is fixed, and the portion where the fluorescent material emits fluorescence is irradiated (incident) with the excitation light. It is moved (shifted) away from the light source over time. Then, the intensity of the fluorescence from the moved fluorescent site is converted into an electric signal using a photodiode, and the amount of the analyte component is measured.
  • the size of the indicator layer 5 for such measurement is designed based on the fluorescent substance concentration, the molecular extinction coefficient at the excitation light wavelength of the fluorescent substance, the signal intensity to be obtained, the usage time, and the shape and size of the fluorescent sensor 1.
  • the size of the indicator layer 5 is such that when the molecular extinction coefficient of the fluorescent material is 3000 to 5000 / M (mol) / cm, the fluorescent material (monomer ) Concentration of 1 to 20%, the length of the indicator layer 5 (that is, the distance away from the light source 6 and the excitation light path length L in FIG. 1A) is 0.1 mm to 3 mm, the indicator layer 5 (that is, the width in the direction orthogonal to the length, and the width W in FIG. 1B) is preferably 10 ⁇ m to 3 mm, and the thickness of the indicator layer 5 is preferably 10 ⁇ m to 500 ⁇ m.
  • the reason why the molecular extinction coefficient of the fluorescent substance is assumed to be 3000 to 5000 / M (mol) / cm is that the fluorescent substance in the case of measuring glucose as an analyte is assumed.
  • the concentration of the fluorescent substance of 1 to 20% is also the amount of the fluorescent substance usually used when measuring glucose.
  • the length of the indicator layer 5 is less than 0.1 mm, all of it deteriorates in the initial stage of irradiation with excitation light, and there is a possibility that the lifespan cannot be extended.
  • such a length may be further shortened depending on the energy intensity of the excitation light, the irradiation time, and the desired lifetime. For example, a functional length of 10 ⁇ m or more is sufficient.
  • the width and thickness may be larger as long as the size of the entire fluorescent sensor 1 is acceptable.
  • the excitation light reaching the farthest position from the light source 6 in the indicator layer 5 is about 1% of the light source exit.
  • the deterioration rate of the fluorescent material depends on the amount of excitation light and the irradiation time. For example, if the fluorescence is continuously irradiated with excitation light of 5 mW / cm 2 , the fluorescence decreases at a rate of 0.8% / min. On the other hand, the absorbance of the excitation light decreases at 0.4% / min. Assuming that the light source 6 is a parallel light beam, it deteriorates at this speed in the immediate vicinity of the light source 6, but the speed is 1/10 at a position 0.5 mm away from the light source and 1/100 at a position 1 mm away. As the deterioration progresses, the absorption of the excitation light near the light source 6 decreases. Therefore, light reaches a farther position in the indicator layer 5, and the portion that emits fluorescence most efficiently shifts to a portion far from the light source 6.
  • fluorescence at 3 mW / cm 2 and the irradiation time of 0.7% of the rate per minute applying an excitation light to the indicator layer 5 is reduced, while the absorbance of the excitation light irradiation time half Decrease at 0.35% per minute.
  • the rate of decrease in absorbance is small for the rate of decrease in fluorescence.
  • the fluorescence signal weakened by the deterioration can be made stronger.
  • increasing the amount of excitation light but the signal strength may be increased, resulting in accelerating the deterioration as described above (the case of the excitation light 5mW / cm 2).
  • the excitation light is irradiated in the longitudinal direction of the indicator layer 5, the fluorescence emission portion is shifted in accordance with the deterioration, so that it can be used for a long time even if the excitation light is strengthened.
  • the excitation light energy is weakened, the degradation rate of the fluorescent substance itself is kept low, the excitation light energy is raised when the signal weakens, and then the strong energy excitation By using light, it can be used longer than applying strong energy from the beginning.
  • the analyte is glucose contained in the interstitial fluid
  • it can be obtained as follows.
  • the current value of the photodiode and the glucose concentration have the following relationship (1).
  • the glucose concentration is calculated based on the concentration of the glucose / fluorescent substance conjugate. Therefore, when the concentration of the fluorescent substance capable of emitting fluorescence is lowered due to deterioration, the concentration of the conjugate of glucose and the fluorescent substance is lowered even at the same glucose concentration.
  • the conjugate of the fluorescent substance and glucose emits fluorescence and becomes a current signal of the photodiode 3.
  • the amount of the fluorescent substance is reduced due to deterioration, the conjugate of the fluorescent substance and glucose is reduced even at the same glucose concentration. As the concentration decreases, the photodiode current decreases.
  • the calibration obtains a photodiode current value obtained when the fluorescent sensor 1 is brought into contact with a known measurement object of glucose concentration, and corrects the coefficient A in the above-described equation (1).
  • B is a bias (a value that does not depend on the concentration of the fluorescent substance-glucose conjugate), correction depending on the glucose concentration is not necessary. However, since it depends on the amount of excitation light, when the energy of the excitation light is changed, it is also necessary to obtain this corresponding to the energy.
  • This type of correction can be performed with higher accuracy by measuring and setting in advance how much excitation light is irradiated (irradiation time x irradiation energy) and how much deterioration occurs in the device. It can be performed.
  • the analyte is glucose
  • the calibration can be similarly performed when measuring other substances.
  • analyte components include, for example, biological components such as lactic acid.
  • the measurement target is not limited to the components in the living body, and can be used as a sensor for indicating a physiological state such as a pH value.
  • FIG. 6 is an external perspective view of the sensor system
  • FIG. 7 is a schematic view when a guide needle of the sensor system is inserted into a living body.
  • the sensor system 100 includes a fluorescent sensor 1 that is inserted and embedded in a living body 200, and an electronic circuit unit 101 that measures and analyzes an analyte component detected by the fluorescent sensor 1 based on a signal from the fluorescent sensor 1. .
  • the fluorescence sensor 1 is embedded at the tip of a guide needle 102 stabbed into the living body 200 (see FIG. 7).
  • the electronic circuit unit 101 is accommodated in, for example, a resin case (the appearance shown is a resin case.
  • the electronic circuit unit 101 contained in the resin case measures a signal from the fluorescent sensor 1.
  • a battery is built in.
  • the electronic circuit unit 101 (in the resin case) has a built-in device for receiving electrodes from the outside in a non-contact manner such as electromagnetic induction without incorporating the battery You may do it.
  • FIG. 8 is an enlarged perspective view of the guide needle.
  • a general stainless steel puncture needle used for medical purposes in which a slit 103 is formed is used.
  • Fluorescence sensor 1 is arranged inside the tip of guide needle 102.
  • the fluorescent sensor 1 is arranged so that the surface with the light shielding film 7 faces the direction of the slit 103. As a result, the analyte in the blood that has entered the guide needle 102 through the slit 103 easily passes through the light shielding film 7 and enters the indicator layer 5 of the fluorescent sensor 1.
  • the electrical connection between the fluorescent sensor 1 and the electronic circuit is performed by wiring through the guide needle 102.
  • the metal wires of 1 ⁇ m to several tens of ⁇ m may be arranged in the guide needle 102 after being insulated and connected to the fluorescent sensor-like wires.
  • the optical fiber 68 is used for the light source 6, the optical fiber 68 is installed together with the wiring from the photodiode 3 according to the inner diameter of the guide needle 102.
  • the inner diameter of the guide needle is about several hundreds of micrometers, the thickness of the optical fiber 68 is slightly smaller than that.
  • FIG. 9 is a schematic cross-sectional view for explaining a modification of the fluorescent sensor.
  • the light from the light source is uniformly irradiated in the parallel direction to the indicator layer as parallel light, but actually, the light is emitted toward the indicator layer as diffused light spreading at the exit of the light source.
  • the indicator layer light is weakened by absorption. This is because even if the fluorescent substance in the indicator layer deteriorates and light can easily pass therethrough, it does not become completely transparent, so that excitation light is weakened by passing therethrough. For this reason, when the position which emits fluorescence most shifts, the excitation light reaching the portion is weaker than the portion immediately before the light source, and as a result, the fluorescence signal becomes small.
  • this modification has a structure that enables correction to increase the light quantity of the light source.
  • a plurality of photodiodes 301 to 306 are provided in the longitudinal direction of the indicator layer 5 (the direction of the excitation light emitted from the light source), and among the photodiodes 301 closest to the light source 6, The optical filter is not provided.
  • the output signal of the photodiode 301 closest to the light source 6 and the output signals of the respective photodiodes 302 to 306 located under the optical filter are compared with time.
  • the output signals of the photodiodes 302 to 306 at the positions change with time with respect to the output signal of the photodiode 301.
  • the change corresponds to a position where the fluorescent substance is deteriorated and the position where the fluorescence is emitted is shifted.
  • the reason why the distance between the photodiodes 301 and 302 is larger than the distance between the photodiodes 302 to 306 is that there is no optical filter 4 on the photodiode 301 and light from the light source 6 enters directly. This is to prevent the influence from adversely affecting the photodiode 302.
  • the fluorescent sensor shown in FIG. 9 As a specific form of use, using the fluorescent sensor shown in FIG. 9, as with calibration, using an analyte having a known concentration while changing with time, the sum of the output signals of the photodiodes 302 to 306 is used.
  • the output signal of the photodiode 301 when the intensity of the light source 6 is changed so as to be the same is measured and recorded along with the elapsed time.
  • a light source is used so that the output signal of the photodiode 301 is recorded corresponding to the measurement time (excitation light irradiation time). If the energy of the excitation light from is changed, accurate measurement can be performed without performing calibration each time.
  • the fluorescence sensor shown in FIG. 9 is used once to change the intensity of the light source 6 so that the output signals of the photodiodes 302 to 306 become the same. If the change with time of the output signal is recorded, the fluorescence sensor having the same magnitude, the same composition and the same concentration of the fluorescent substance indicator layer can be applied and corrected in the same manner. For this reason, for example, as shown in FIG. 10, the photodiode under the optical filter 4 is applicable even if it is not divided. In FIG. 10, a photodiode 301 closest to the light source 6 and one photodiode 307 located under the optical filter 4 are provided. That is, FIGS. 9 and 10 have at least two photodiodes for individually measuring the received light amount, and the photodiode 301 closest to the light source 6 does not have the optical filter 4.
  • the present embodiment absorbs the optical characteristics of the fluorescent substance in the indicator layer, that is, the light of the excitation light, is transparent to the light of the fluorescence wavelength, and deteriorates when the excitation light is applied. Focusing on the characteristic that the ability to absorb excitation light at the same time as the energy decreases, the excitation light is irradiated toward the indicator layer in parallel with the photodiode light receiving surface. As a result, the fluorescent material in the indicator layer gradually deteriorates from the light source toward the back and becomes transparent, and the position where the fluorescence is emitted is shifted most. As a result, the measurement can be continued until the excitation light reaches the innermost point from the light source of the indicator layer, and the lifetime is longer than when the excitation light is applied to the surface of the indicator layer as in the prior art. can do.
  • the fluorescent sensor of the present invention is not limited to such a usage pattern and can be used in various forms.
  • the fluorescence sensor and the electronic circuit can be formed on the same substrate or placed in the same package and used for measuring glucose concentration and the like.
  • measurement is performed by dropping a liquid to be measured on the indicator layer. In this case, it is not necessary to provide a light shielding film by placing the indicator layer of the fluorescent sensor in a light shielding case.
  • Fluorescent sensor 1 Fluorescent sensor, 2 substrates, 3 photodiode, 4 optical filters, 5 indicator layer, 6 Light source, 7 light shielding film, 61 LED, 62 Prism, 63 angle filter, 64 wiring layers, 65 pedestal, 66 faces 68 Optical fiber, 100 sensor system, 101 electronic circuit part, 102 guide needle, 103 slit.

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  • Health & Medical Sciences (AREA)
  • Immunology (AREA)
  • Chemical & Material Sciences (AREA)
  • Physics & Mathematics (AREA)
  • Analytical Chemistry (AREA)
  • Optics & Photonics (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Pathology (AREA)
  • Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
  • Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
  • Measurement Of The Respiration, Hearing Ability, Form, And Blood Characteristics Of Living Organisms (AREA)

Abstract

La présente invention porte sur un capteur de fluorescence capable de prolonger la durée de vie d'une couche d'indicateur. Le capteur de fluorescence est caractérisé en ce qu'il a une couche d'indicateur (5) pour détecter un composant d'analyte d'un organisme, la couche comprenant une substance fluorescente qui émet une fluorescence selon la quantité du composant d'analyte par exposition à une lumière d'excitation, une photodiode (3) pour recevoir la fluorescence et délivrer en sortie un signal électrique selon l'intensité de la fluorescence, et une source lumineuse (6) pour rayonner la lumière d'excitation sur la couche d'indicateur (5). La couche d'indicateur (5) est agencée sur la photodiode (3) et la lumière d'excitation émise par la source lumineuse (6) est irradiée sur la couche d'indicateur (5) dans une direction parallèle à la surface de réception de lumière de la photodiode (3).
PCT/JP2012/073195 2011-09-12 2012-09-11 Capteur de fluorescence et procédé de mesure de composant d'analyte Ceased WO2013039065A1 (fr)

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Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH0643965B2 (ja) * 1986-04-23 1994-06-08 ア−・フアウ・エル ア−・ゲ− 物質濃度を測定するためのセンサ素子
JP2000146944A (ja) * 1998-11-16 2000-05-26 Sakura Color Prod Corp ガス検知装置及びガス検知方法
JP2001340072A (ja) * 2000-03-29 2001-12-11 Matsushita Seiko Co Ltd 微生物計量装置
JP2001525930A (ja) * 1997-05-13 2001-12-11 コルビン，アーサー・イー・ジュニア 改良された蛍光検出デバイス
WO2005095929A1 (fr) * 2004-03-30 2005-10-13 Hamamatsu Photonics K.K. Élément de masquage, méthode de mesure de la lumière, kit de mesure de la lumière et conteneur de mesure de la lumière

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH0643965B2 (ja) * 1986-04-23 1994-06-08 ア−・フアウ・エル ア−・ゲ− 物質濃度を測定するためのセンサ素子
JP2001525930A (ja) * 1997-05-13 2001-12-11 コルビン，アーサー・イー・ジュニア 改良された蛍光検出デバイス
JP2000146944A (ja) * 1998-11-16 2000-05-26 Sakura Color Prod Corp ガス検知装置及びガス検知方法
JP2001340072A (ja) * 2000-03-29 2001-12-11 Matsushita Seiko Co Ltd 微生物計量装置
WO2005095929A1 (fr) * 2004-03-30 2005-10-13 Hamamatsu Photonics K.K. Élément de masquage, méthode de mesure de la lumière, kit de mesure de la lumière et conteneur de mesure de la lumière

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