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WO2013161991A1 - Fluorescence sensor - Google Patents

Fluorescence sensor

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Publication number
WO2013161991A1
WO2013161991A1 PCT/JP2013/062368 JP2013062368W WO2013161991A1 WO 2013161991 A1 WO2013161991 A1 WO 2013161991A1 JP 2013062368 W JP2013062368 W JP 2013062368W WO 2013161991 A1 WO2013161991 A1 WO 2013161991A1
Authority
WO
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Application
Patent type
Prior art keywords
sensor
fluorescence
light
indicator
portion
Prior art date
Application number
PCT/JP2013/062368
Other languages
French (fr)
Japanese (ja)
Inventor
亮 太田
Original Assignee
オリンパス株式会社
テルモ株式会社
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date

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    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61BDIAGNOSIS; SURGERY; IDENTIFICATION
    • A61B5/00Detecting, measuring or recording for diagnostic purposes; Identification of persons
    • A61B5/145Measuring characteristics of blood in vivo, e.g. gas concentration, pH value; Measuring characteristics of body fluids or tissues, e.g. interstitial fluid, cerebral tissue
    • A61B5/14503Measuring characteristics of blood in vivo, e.g. gas concentration, pH value; Measuring characteristics of body fluids or tissues, e.g. interstitial fluid, cerebral tissue invasive, e.g. introduced into the body by a catheter or needle or using implanted sensors
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61BDIAGNOSIS; SURGERY; IDENTIFICATION
    • A61B5/00Detecting, measuring or recording for diagnostic purposes; Identification of persons
    • A61B5/0059Detecting, measuring or recording for diagnostic purposes; Identification of persons using light, e.g. diagnosis by transillumination, diascopy, fluorescence
    • A61B5/0071Detecting, measuring or recording for diagnostic purposes; Identification of persons using light, e.g. diagnosis by transillumination, diascopy, fluorescence by measuring fluorescence emission
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61BDIAGNOSIS; SURGERY; IDENTIFICATION
    • A61B5/00Detecting, measuring or recording for diagnostic purposes; Identification of persons
    • A61B5/145Measuring characteristics of blood in vivo, e.g. gas concentration, pH value; Measuring characteristics of body fluids or tissues, e.g. interstitial fluid, cerebral tissue
    • A61B5/14532Measuring characteristics of blood in vivo, e.g. gas concentration, pH value; Measuring characteristics of body fluids or tissues, e.g. interstitial fluid, cerebral tissue for measuring glucose, e.g. by tissue impedance measurement
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00Investigating or analysing materials by the use of optical means, i.e. using infra-red, visible or ultra-violet light
    • G01N21/62Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64Fluorescence; Phosphorescence
    • G01N21/6428Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00Investigating or analysing materials by the use of optical means, i.e. using infra-red, visible or ultra-violet light
    • G01N21/62Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64Fluorescence; Phosphorescence
    • G01N21/645Specially adapted constructive features of fluorimeters
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61BDIAGNOSIS; SURGERY; IDENTIFICATION
    • A61B2562/00Details of sensors; Constructional details of sensor housings or probes; Accessories for sensors
    • A61B2562/12Manufacturing methods specially adapted for producing sensors for in-vivo measurements
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00Investigating or analysing materials by the use of optical means, i.e. using infra-red, visible or ultra-violet light
    • G01N21/62Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64Fluorescence; Phosphorescence
    • G01N2021/6417Spectrofluorimetric devices
    • G01N2021/6421Measuring at two or more wavelengths
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00Investigating or analysing materials by the use of optical means, i.e. using infra-red, visible or ultra-violet light
    • G01N21/62Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64Fluorescence; Phosphorescence
    • G01N21/6428Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
    • G01N2021/6439Measuring 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
    • G01N2021/6441Measuring 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 with two or more labels
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00Investigating or analysing materials by the use of optical means, i.e. using infra-red, visible or ultra-violet light
    • G01N21/75Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
    • G01N21/77Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator
    • G01N2021/7769Measurement method of reaction-produced change in sensor
    • G01N2021/7786Fluorescence
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00Investigating or analysing materials by the use of optical means, i.e. using infra-red, visible or ultra-violet light
    • G01N21/75Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
    • G01N21/77Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator
    • G01N21/78Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator producing a change of colour
    • G01N21/80Indicating pH value

Abstract

A fluorescence sensor (4) is provided with: a substrate part (20) in which a plurality of recesses (23A, 23B) are formed in a principal surface (20SA), PD elements (12A, 12B) for receiving fluorescence and outputting a detection signal being formed on wall surfaces of the recesses (23A, 23B), respectively; filters (14A, 14B) for covering the PD elements (12A, 12B) and blocking excitation light; indicators (17A, 17B) for generating fluorescence in accordance with the quantity of an analyte when excitation light is received, the indicators (17A, 17B) being provided inside the recesses (23A, 23B), respectively; a light-shielding layer (18) for covering openings of the recesses (23A, 23B) and blocking the penetration of external light into the indicators (17A, 17B) but passing an analyte (9); and LEDs (15A, 15B) for irradiating excitation light to the indicators (17A, 17B), respectively, from a bottom side of the recesses (23A, 23B).

Description

Fluorescence sensor

The present invention relates to a fluorescence sensor for measuring the concentration of analyte, for fluorescent sensor is particularly small fluorescence spectrophotometer manufactured using semiconductor manufacturing technology and MEMS technology.

Existence confirmation of the analyte i.e. the measurement substance in a liquid, or a variety of analyzers for measuring the concentration has been developed. For example, the transparent container of constant volume, a fluorescent dye that generates a property changes fluorescence by the presence of the analyte, was injected and the measured solution containing analyte, the fluorescence intensity from the irradiated fluorochrome excitation light fluorescence spectrophotometer that measures the analyte concentration is known by measuring.

Small fluorescence spectrophotometer includes an indicator containing a light detector and the fluorescent dye. Then, when the analyte to be measured solution is irradiated with excitation light from a light source to possible ingress indicator, a fluorescent dye in the indicator generates the fluorescence amount corresponding to the analyte concentration of the measured solution, the fluorescent light detector receives light. Photodetector is a photoelectric conversion element, and outputs an electric signal corresponding to the amount of light received. Is the analyte concentration of the measured solution from the electrical signal is measured.

Recently, in order to measure the analyte traces in the sample, small fluorescent spectrophotometer has been proposed which is manufactured by using semiconductor manufacturing technology and MEMS technology. Hereinafter, the micro fluorometers, referred to as "fluorescence sensor".

For example, a fluorescent sensor 110 shown in FIGS. 1 and 2 is disclosed in U.S. Patent No. 5,039,490. Fluorescence sensor 110, the excitation light is capable of transmitting the transparent supporting substrate 101, a photoelectric conversion element 103 that converts the fluorescence into an electric signal, an optical plate 105 having a light collecting function unit 105A for condensing the excitation light , an indicator 117 which generates fluorescence upon incidence of excitation light by interacting with the analyte 9, a cover layer 109, and a.

The photoelectric conversion element 103 is formed on a substrate 103A made of, for example, silicon. Substrate 103A is not transmitted through the excitation light. Therefore, the fluorescence sensor 110, the excitation light has a permeable void region 120 around the photoelectric conversion element 103.

That is, only the excitation light E which passes through the gap region 120, by the action of light collecting function unit 105A, in the indicator 117, and is focused near the top of the photoelectric conversion element 103. A focused excitation light E, by the interaction of the analyte 9 which enters the interior of the indicator 117, the fluorescence F is generated. Some of the generated fluorescence is incident on the photoelectric conversion element 103, the fluorescence intensity in the photoelectric conversion element 103, i.e. the signal, such as current or voltage proportional to the concentration of analyte 9 occurs. Note excitation light, by the action of the filter formed on the photoelectric conversion element 103 (not shown), not incident on the photoelectric conversion element 103.

As explained above, the fluorescence sensor 110, a photo-diode which is a photoelectric conversion element 103 to the substrate 103A that is stacked on the transparent supporting substrate 101, on which, an optical plate 105 and the indicator 117 It is stacked.

However, the above fluorescent sensor 110, and a region of the void region 120 and the photoelectric conversion element 103 is a path of the excitation light, having on the same plane. Thus, when large area of ​​the void region 120 is a passage for guiding more excitation light indicator 117, the area of ​​the photoelectric conversion element 103 is narrowed, not to increase the sensitivity of the fluorescence sensor . A broad area of ​​the photoelectric conversion element 103 in order to increase the detection sensitivity of the photoelectric conversion element 103 in the opposite, excitation light area of ​​the void region 120 is a passage of the excitation light is guided to become the indicator 117 narrow decreases to put away, not also to increasing the sensitivity of the fluorescence sensor. That is, in the fluorescent sensor 110, to obtain a high detection sensitivity is not easy.

The present invention aims to provide a high detection sensitivity fluorescence sensor.

Fluorescence sensor of one embodiment of the present invention, a plurality of recesses are formed on the main surface, and a substrate portion in which the photoelectric conversion element that outputs a detection signal received fluorescence to the wall of each recess is formed, the covering the photoelectric conversion element, and a filter that blocks excitation light, is disposed within each of said recesses, when receiving the excitation light and the indicator for generating the fluorescence corresponding to the analyte amount, of the recess to cover the opening, but to block the external light entering into the indicator, the analyte is provided with a light shielding layer that passes through, and a light emitting element for irradiating excitation light to the indicator from the bottom side of the recess.

It is a cross-sectional view of a conventional fluorescent sensors. Is an exploded view illustrating the structure of a conventional fluorescence sensor. It is an explanatory view for explaining a sensor system having a fluorescence sensor of the first embodiment. Is an exploded view illustrating the structure of the fluorescence sensor of the first embodiment. It is a cross-sectional view of a fluorescent sensor of the first embodiment. It is a sectional view for explaining a manufacturing method of a fluorescent sensor of the first embodiment. It is a sectional view for explaining a manufacturing method of a fluorescent sensor of the first embodiment. It is a sectional view for explaining a manufacturing method of a fluorescent sensor of the first embodiment. It is a sectional view for explaining a manufacturing method of a fluorescent sensor of the first embodiment. It is a sectional view for explaining a manufacturing method of a fluorescent sensor of the first embodiment. Is an exploded sectional view illustrating the structure of the fluorescence sensor of the second embodiment. Is an exploded sectional view illustrating the structure of the fluorescence sensor of the third embodiment. It is a sectional view for explaining a manufacturing method of a fluorescent sensor of the fourth embodiment. It is a top view for explaining an arrangement of the indicator fluorescence sensor of the first modification. It is a top view for explaining an arrangement of the indicator fluorescence sensor of the first modification. It is a top view for explaining an arrangement of the indicator fluorescence sensor of the first modification. It is a top view for explaining an arrangement of the indicator fluorescence sensor of the first modification. It is a cross-sectional view for explaining a fluorescent sensor of the second modification. It is a cross-sectional view for explaining a fluorescent sensor of a modification 3. It is a cross-sectional view for explaining a fluorescent sensor of a modification 4.

<First Embodiment>
The following describes fluorescence sensor 4 of the first embodiment of the present invention.

As shown in FIG. 3, the fluorescence sensor 4 includes a main body 2 and receiver 3, together constitute the sensor system 1. That is, the sensor system 1 includes a fluorescent sensor 4, the main body portion 2, a receiver 3 for storing the received signals from the main unit 2, a. Transmission and reception of signals between the main body portion 2 and receiver 3 is performed in a wireless or wired.

Fluorosensor 4 of the needle structure includes a needle portion 7 having a needle tip portion 5 and an elongated needle body portion 6 having a sensor unit 10 which is a main function unit, a connector portion which is integrated with the rear end portion of the probe main body portion 6 8, comprises a. Needle tip 5, the needle body 6, and the connector portion 8 may be integrally formed of the same material.

Connector portion 8 is fitted detachably fitting portion 2A of the main body portion 2. A plurality of wires 60 extending from the sensor portion 10 of the fluorescence sensor 4 by connectors 8 are fitted portion 2A and mechanically engagement of the main body portion 2 is electrically connected to the main body 2 .

Body 2 has a controller 2B which a CPU or the like for a driving and control of the sensor unit 10, an operation unit 2C including a CPU or the like for processing the signal output from the sensor unit 10, a. At least one of the control unit 2B or the arithmetic unit 2C is, may be disposed in the connector portion 8, and the like of the fluorescent sensor 4 may be arranged on the receiver 3. The control unit 2B and a calculation unit 2C may be the same CPU.

In the case of the wired transmitting and receiving between the receiver 3, the body portion 2 has a signal line instead of the radio antenna. Also, if having a memory portion of the required capacity body 2 receivers 3 is not required.

Fluorescence sensor 4 in a state fitted to the body portion 2, the subject himself needle tip 5 to puncture the body surface is indwelling. Then, for example, the glucose concentration in the body fluid is measured continuously and stored in a memory of the receiver 3. That is, the fluorescence sensor 4 is a needle-type sensor for measuring the body analyte is a short subcutaneous indwelling of about one week of continuous use period. However, not insert fluorescence sensor 4 into the body, collected body fluid or a body fluid circulated body through the flow path outside the body, it may be contacted in vitro.

Then, as shown in FIGS. 4 and 5, the sensor unit 10 is a main functional unit of the fluorescence sensor 4 includes a substrate 20, a light-emitting element generating excitation light-emitting diode (Light Emitting Diode: the "LED It has also referred to) 15A, and 15B and "indicator 17A for generating a fluorescence corresponding to the excitation light and the analyte quantity, and 17B, and the light shielding layer 18, a. In the fluorescent sensor 4, by the light shielding layer 18 is in contact with blood or body fluid in vivo, the analyte 9 enters the indicator 17 through the light shielding layer 18.

Substrate 20 includes a wiring board unit 30 is a first substrate portion, a second substrate portion in which two through-holes 46A to the principal surface 20SA (principal surface 20SB), 46B is formed frame-like substrate portion are produced by joining 40, the via the adhesive layer 13. Therefore, the substrate 20 is a multi-cavity with two recesses 23A, 23B in the main surface 20SA. That is, the surface of the wiring substrate section 30 is a recess 23A, 23B bottom surface of the through hole 46A of the frame-shaped substrate 40, the wall surfaces of the 46B, the recess 23A, a wall of 23B.

Incidentally, for example, a substrate portion 20 made of N-type semiconductor, i.e. appropriately in the surface or the like of the wiring board 30 and the frame-like substrate portion 40, the insulating layer is formed is not shown. In the following, when referring to a respective component of the same function is omitted trailing character (A or B). For example, the through holes 46A, respectively 46B of the through-hole 46.

Inside the recess 23A, the indicator 17A and has a is disposed LEDs 15A, and the indicator 17B and LED15B are disposed inside the recess 23B. The light emitting element is not limited the LED. However, the light generation efficiency, the wavelength selectivity of the breadth of the excitation light, and light of wavelength other than ultraviolet as the excitation light from the viewpoint of not only occur slightly, substantially rectangular chip-like LED is preferred.

Indicator 17 generates a fluorescent light amount corresponding to the amount of analyte 9 by interaction and excitation light with the analyte 9 having entered. For example, the indicator 17 to the excitation light of wavelength 375 nm, to generate, for example, wavelength 460nm fluorescence of longer wavelength. The thickness of the indicator 17 is set to about several tens of μm ~ 200μm. Indicator 17 is configured amount of analyte 9, i.e. from a base material that contains a fluorescent dye that emits fluorescence having an intensity corresponding to the analyte concentration in the sample.

Fluorescent dyes are selected according to the type of analyte 9, the amount of fluorescence generated in accordance with the amount of analyte 9 if reversibly changing fluorescent dye, can be used to look like. That is, the fluorescence sensor 4, by the choice of the fluorescent dye, an oxygen sensor, a glucose sensor, pH sensor, such as an immunosensor or a microorganism sensor, corresponds to the variety of applications.

Indicator 17, for example, the water easily hydrogel as the base material, it is contained to or bind the fluorescent dye in the hydrogel. Polysaccharides such as cellulose or dextran as a component of the hydrogel is prepared from (meth) acrylamide, methylol acrylamide or inhibit dollars carboxymethyl acrylic hydrogels are prepared by polymerizing a monomer such as ethyl acrylate, or polyethylene glycol with a diisocyanate, urethane-based hydrogels can be used.

Indicator 17 may be joined to LED15 through a transparent protective layer for protecting the LED15. The transparent protective layer, a silicon oxide, an epoxy resin, a silicone resin, or a transparent amorphous fluorocarbon resin, can be used. Transparent protective layer to have electrical insulating properties, having a moisture barrier properties, to have good transmittance for the excitation light and fluorescence, is selected from materials having characteristics such as. As properties of the transparent protective layer, it is important occurrence of fluorescence excitation light in the layer be irradiated is small. The characteristic of this fluorescence is small, it is needless to say important property of all transparent material of the fluorescent sensor 4 except the indicator.

Light-shielding layer 18 is formed on the upper surface side of the indicator 17, are the following layers a few tens of μm thick. Incidentally, the light blocking layer 18 may be divided between a use indicator 17A and an indicator 17B.

On the other hand, the wall surface of the through-holes 46A of the frame-shaped substrate 40, i.e. on the wall surface of the concave portion 23A of the substrate portion 20, a photodiode which is a photoelectric conversion element that outputs a detection signal received fluorescence (Photo Diode: hereinafter "PD" that also referred) element 12A is formed similarly PD element 12B also on the wall surface of the through hole 46B is formed. That is, the light receiving portion 12D of the PD element 12 is provided so as to surround the indicator 17 is formed so as the light receiving surface faces the indicator 17. Hereinafter, the light receiving portion 12D of the PD element 12.

PD element 12 may be formed on the entire wall surface but, in order to receive only the fluorescence efficiently, may be formed only in the region opposed to the indicator 17. The PD element 12 may be formed on all the wall surfaces in four sides, or may be formed only on a part of the surface.

That, PD element 12 may be formed on at least part of the wall of the recess 23. As the photoelectric conversion element, photoconductor (photoconductor), or phototransistor (Photo Transistor, PT) or the like.

Formed on the wall surface, each of the PD elements 12A, filter 14A to cover 12B, 14B are disposed. PD elements 12A, filter 14 covering at least a portion of the 12B is to cut off the excitation light, the fluorescence of longer wavelength than the excitation light is high pass type absorption filter of passing. As the material of such a filter, a silicon layer or a silicon carbide layer is preferable. For example, the silicon layer and the silicon carbide layer, whereas the 375nm In the transmittance of the excitation light wavelength is 10 -5% or less, and the transmittance of 10% or more 460nm emission wavelengths (transmittance of the excitation light wavelength / having 6 or more digits transmittance selectivity as the ratio of the fluorescence transmission wavelengths). The transmittance of the fluorescence wavelength may be a high band-pass filter as the filter 14. Further, the transparent layer that protects the PD element 12, for example may be filter 14 is provided through the silicon oxide layer.

Wiring board 30, three wires 51A for transmitting the detection signal from the PD elements 12A, 12B, 52,51B, and LEDs 15A, a wiring layer consisting of wiring 53 and 54 for transmitting a driving signal to 15B with 50. Wires 52, N-type semiconductor such as frame-like substrate portion 40 partially N-type impurity into the surface of, for example, by introducing phosphorus or arsenic, formed with higher conductivity low-resistance region 12H, is connected there. Incidentally, LEDs 15A, when driving 15B separately, each wire is arranged. Further, the wiring 51 and the wiring 51B may be connected to any of a plurality of wires 60 for transmitting a signal to the main body portion 2 from being connected.

Material of the frame-shaped substrate 40 is single crystal silicon is preferable in order to form the PD element 12 to the frame-shaped substrate 40 may be a glass or ceramic. In that case, the surface and the frame surface of one 20SB of the main surface of the substrate portion 40 of the recess 23, a semiconductor thin film such as the following polysilicon or amorphous silicon a few microns is formed, it PD element 12 is formed.

Excitation light LED15 occurs, are irradiated to the fluorescent dye in the indicator 17. Then, a part of the fluorescence fluorescent dye is generated by interaction between the analyte 9, and reaches the PD element 12 through the filter 14, it is converted into the detection signal.

Fluorescence sensor 4 to detect fluorescence by PD element 12 formed on a wall surface surrounding the indicator 17, the detection sensitivity is high.

Furthermore, the fluorescent sensor 4, two recesses 23A, PD elements 12A respectively on the wall of 23B, 12B are formed. Therefore, a wide area of ​​the light receiving portion 12D of the wall area i.e. PD element 12. Furthermore, fluorescence fluorescent dyes because of the short distance to the light receiving portion is generated in the PD element 12 from the center of the recess 23 reaches the PD element 12 without being attenuated. Therefore, the fluorescence sensor 4, the signal output is high, for a good wide dynamic range of S / N ratio, the detection sensitivity is high.

Next, brief description will be given of a manufacturing method of the fluorescent sensor 4. Although a partial cross-sectional view of a region of FIG. 6A ~ FIG 6E In one fluorescent sensor 4, the actual process, to form a number of the sensor unit 10 collectively as a wafer process.

First, as shown in FIG. 6A, in the production of the frame-shaped substrate 40, the silicon wafer 40W conductivity (N-type), the etching performed through a mask layer 71, a large number of through-holes 46A, 46B is formed. The etching can be used various known methods.

In order the size of the opening of the through hole 46 are designed in accordance with the specifications, distribution 設箇 plants are needle tip 5, a plurality of openings, for example, vertical 150 [mu] m, as the lateral 500μm elongated preferably disposed in the area, one opening is preferably several hundred μm a size of about from 10 [mu] m.

Also, the fluorescent sensor 4, but the wall surface of the through hole 46 is perpendicular to the main surface, as will be described later, wall predetermined angle, i.e., may have a shape with a taper. Recess of tapered shape can be produced, for example, by wet etching.

Next, as shown in FIG. 6B, PD element 12 is formed on the wall surface of the through hole 46. That is, in an inclined state a silicon wafer 40W on which the mask layer 72 is formed on the 5 to 30 degrees, the ion implantation process is performed from four directions. For example, conditions for implanting boron (B), the acceleration voltage: 10 ~ 100 keV, implantation dose: a 1 × 10 15 cm -2 order.

Then, on the PD element 12 of the wall surface of the through hole 46 of the silicon wafer 40W, filter 14 by the CVD method, is formed.

Separately, a silicon wafer 30W conductivity comprising a wiring board unit 30 (N-type) is prepared. The silicon wafer 30 W, although not shown, after the insulating film such as an oxide film is formed, the wiring 51,52,51B and LED15A for transmitting a detection signal from the PD element 12, a drive signal to 15B wires 53 and 54 for supplying is formed by a sputtering method or an evaporation method, or the like.

As shown in FIG. 6C, a silicon wafer 40W is upside down, the main surface 20SB of the silicon wafer 40W are bonded to the silicon wafer 30W via the adhesive layer 13.

As shown in FIG. 6D, the two wafers bonded are joined wafer 20W, the through-hole 46 of the frame-shaped substrate portion 40 becomes a recess 23 with a bottom. Then the recess 23A of the junction wafer 20W, the bottom surface of each of 23B, LEDs 15A, 15B are disposed. The arrangement of LED 15, bonding method using such an optically transparent acrylic resin or silicone resin, or a method such as various bonding methods such as flip chip bonding method can be used.

Then, as shown in FIG. 6E, the recess 23A of the bonding wafer 20W, within each of 23B, the indicator 17A, 17B is arranged.

Next, the light-shielding layer 18 is the indicator 17A, the sensor substrate 10W is formed on the 17B is completed. The light-shielding layer 18 is made of a pore structure of submicron size, metal, inorganic thin film or the ceramics, composite structure of the hydrogel such that the carbon black is mixed in the base material of organic polymer such as polyimide or polyurethane or, , the resin was mixed with analyte permeable polymer to the carbon black, such as celluloses or polyacrylamides, or they use a laminated resin or the like.

By the sensor substrate 10W is singulated, a number of the sensor portion 10 is manufactured at once. The sensor unit 10 has been produced separately, by being joined to the front end portion of the probe main body portion 6 which extends from the connector portion 8, the fluorescence sensor 4 is completed.

As a method for producing a fluorescent sensor, distribution is not limited to this, and the wiring board portion 30 which is sectioned, after joining the frame-shaped board portion 40 which is sectioned, into recesses 23 LED 15 or the like methods such as to set may be used.

Further, to the first silicon wafer may be processed as extended portion of the wiring board 30 constitute a probe main body portion 6 of the needle 7, a needle body portion 6 which is separately prepared, the fluorescence sensor 4 and a needle tip 5 may constitute a needle portion 7 are joined with.

As explained above, the fluorescence sensor 4 may collectively mass-produced by a wafer process. Therefore, the fluorescence sensor 4 can provide an inexpensive stable quality.

Next, an example of the operation of the fluorescence sensor 4.

LED15 is the center wavelength in a single interval to pulse emission of the excitation light of about 375nm, for example 30 seconds. For example, a pulse current to LED15 is 1 mA ~ 100 mA, the pulse width of the emission is 101 ms ~ 100 ms.

Excitation light LED15 occurs is incident on the indicator 17. Indicator 17 emits fluorescence intensity corresponding to the amount of analyte 9. Incidentally, the analyte 9 passes through the light shielding layer 18, to and from the indicator 17.

Some of the fluorescence indicator 17 occurs through the filter 14 and enters the PD element 12. The fluorescence, by generating the photo-electric conversion by the photo-generated charges in the PD element 12, is outputted as a detection signal. A part also excitation light LED15 occurs, is incident on the wall surface of the recess 23, hardly incident to the PD element 12 by the action of the filter 14.

In the fluorescent sensor 4 performs calculation unit 2C detection signal of the main body portion 2, i.e., the original arithmetic processing the voltage due to photogenerated charge current or accumulated due to light-generated charge from the PD element 12, Ana to calculate the write amount.

Here, the fluorescence sensor 4 has independently two pairs of sensor function unit capable of measuring the analyte, i.e., two pairs of sets of LED15 and PD element 12. Therefore, the fluorescence sensor 4 can achieve the unique effects by a variety of driving methods.

For example, if the deterioration of the indicator by irradiation with excitation light travels may emit light alternately and LED15A and LEDs 15b. Signal output is reduced, but the fluorescence sensor 4 is allowed a longer period of time measured (long life) is.

Moreover, since one of the sensor function portion (LED 15 or the PD element 12 and the like) and the other also failed to continue to measure if operating normally, the fluorescence sensor 4 and the reliability is improved. Also, typically, it was measured using one of the sensor function portion, and the other may be used as a backup.

Further, by changing the cumulative amount of light irradiated to the integrated light amount and LED15B irradiating the LEDs 15A, it may be an indicator 17A and an indicator 17B as a different deterioration states. Since it is possible to correct the fluorescent light amount decreasing due to deterioration, the fluorescence sensor 4 becomes more accurate.

As explained above, the fluorescence sensor 4 having two pairs of sensor function unit, by selecting the control method according to the purpose, has a high output, high reliability, long life, the effect of such high precision.

<Second Embodiment>
It will now be described fluorescence sensor 4A of the second embodiment. Fluorosensor 4A is omitted the description the same reference numerals the same components because of the similar fluorescence sensor 4 of the first embodiment.

As shown in FIG. 7, the fluorescence sensor 4A of the second embodiment, the sensor unit 10A includes a frame-like substrate portion 40A, one wiring board unit 30A which LED15L is mounted in is joined substrate portion 20A It is configured. That is, two concave portions 23A, two indicators 17A filled in 23B, the 17B, 1 single LED15L is irradiated with excitation light from below.

In the fluorescent sensor 4A shown in FIG. 7, the PD element 12A and the PD element 12B are connected, and outputs a detection signal via a common line 51, 52.

Fluorosensor 4A is simpler structure because it one LED15L 2 one indicator 17A, a common excitation light source 17B.

<Third Embodiment>
It will now be described fluorescence sensor 4B according to the third embodiment. Fluorosensor 4B is omitted the description the same reference numerals are given to the same components because of the similar fluorescence sensor 4, 4A.

As shown in FIG. 8, the fluorescence sensor 4B according to the third embodiment, the substrate portion 20B of the sensor unit 10B, of the opening of the through hole 46, the region just below the opposite side of the opening the opening covered with the light shielding layer 18 one LED15L so as to cover is configured to be bonded to the frame-like substrate portion 40B.

That, LED15L has two through holes 46A, covers the region immediately below the opening of 46B principal surface 20SB side. In other words, the through-holes 46A, 46B bottom of which is constituted by the upper surface of LED15L. Incidentally through holes 46A, 46B bottom surface of the can be a transparent adhesive layer 13.

The wiring layer 50 that constitutes a wire connected to the PD elements 12A, 12B and LED15L are arranged in the frame-shaped substrate portion 40B.

Light leakage prevention layer 19 disposed so as to cover the bottom surface (lower surface) and side surfaces of the LED15L, the excitation light emitted from the bottom and sides, and, excited reflected by the second principal surface 20SB of the frame-shaped substrate part 40B light is prevented from light leakage to the outside. That is, has a function of light leakage prevention layer 19 is similar to the light shielding layer 18, the analyte permeability is not required.

As explained above, the fluorescence sensor 4B has the effect of fluorescent sensors 4, 4A, for further configuration is simple, it is easy and more production.

<Fourth Embodiment>
It will now be described fluorescence sensor 4C according to the fourth embodiment. Fluorosensor 4C is omitted the description the same reference numerals are given to the same components because of the similar fluorescence sensor 4.

As shown in FIG. 9, the fluorescence sensor 4C, are integrally fabricated by a silicon substrate portion 20C of the sensor section 10C is a semiconductor. That is, the concave portion 23A, 23B of the substrate portion 20C is a recess formed by etching.

As an etching method, tetramethylammonium hydroxide (TMAH) aqueous solution, dry etching but wet etching and the like aqueous potassium hydroxide (KOH) is desired, reactive ion etching (RIE), etc. chemical dry etching (CDE) law also can be used, for example, in the case of using a substrate made of a silicon (100) plane as the major surface as the silicon is a slow anisotropic etching than the etching rate of the (100) plane of the (111) plane Therefore, the recess 23A, the wall surface of 23B becomes (111) plane, the angle θ1 is 54.74 degrees and (100) plane of the. That is, the wall is tapered.

The recess 23A, the wall surface of 23B, PD elements 12A, respectively, 12B are formed. Recess 23 with a wall taper wall not only is a large area to form a PD device as compared with the vertical recess, forming the PD element 12 is easy.

Incidentally, PD elements 12A, 13A, LEDs 15A, 15B are connected to the wiring layer 50 of the second principal surface 20SB via the through wiring.

As explained above, the fluorescence sensor 4C has the effect of fluorescent sensors 4, for further configuration is simple, it is easy and more production.

<Modification 1>
As shown in FIG. 10A, the fluorescent sensors 4 ~ 4C of the first embodiment, the sensor unit 10 ~ 10C are two plan view (sectional shape) had rectangular recess 23A, the 23B, the concave portion 23 the number, i.e., the number of the indicator 17 may be three or more. For example, it may have four recesses 23A ~ 23D as fluorescent sensors 4D modification shown in FIG. 10B (sensor portion 10D). Further, various shapes such as the cross-sectional shape of the plurality of recesses 23 may be polygonal or elliptical or the like, such as hexagonal, or combinations thereof as fluorescent sensors 4D1 shown in FIG. 10C (sensor portion 10D1) But good. Also as in the fluorescence sensor 4D2 shown in FIG. 10D (sensor portion 10D2), shape and size may combine different recess 23.

The area number is large and the wall surface of the same area of ​​the sensor unit is a also a recess 23 is increased, the fluorescence sensor detection sensitivity becomes higher,
Here, the intensity of the excitation light LED15 occurs is plane distribution. Fluorescence intensity indicator 17 in the region where the excitation light of high intensity is irradiated emits light is high. However, when the excitation light with high intensity is irradiated, the deterioration indicator 17 is likely to be promoted. Fluorescence intensity intensity of the excitation light emitting indicator 17 of lower region incident contrast is low. That is, when there is a large in-plane distribution of the intensity of the excitation light incident on the indicator 17, there is a possibility that the detection sensitivity of the fluorescence sensor is or, or deterioration over time acceleration decreases.

However, it as in the fluorescence sensor 4D2, the opening area of ​​the plurality of recesses 23, the shape and arrangement etc., also averaging the intensity distribution of the excitation light incident on the indicator 17.

Fluorosensor 4D ~ 4D2 variant 1 can be in addition to the effects possessed by the fluorescent sensors 4 ~ 4C, furthermore, not only the more high detection sensitivity, also possible to suppress the deterioration of the indicator 17.

<Modification 2-4>
In the fluorescent sensor 4 ~ 4D2 which has already been described, a plurality of indicators 17 and a plurality of PD elements 12 of the same type of analytes 9, for example, glucose is measured. That is, the fluorescent sensors 4 ~ 4D2 is the selection of the fluorescent dye, an oxygen sensor, a glucose sensor, pH sensor, such as an immunosensor or a microorganism sensor, corresponds to the variety of applications. However, it analytes 9 measurable was one.

In contrast, the sensor portion 10E of the fluorescence sensor 4E of Modification 2 shown in FIG. 11 is a multi-sensing type that can measure a plurality of types of analytes. In fluorosensor 4E, indicator, the first indicator 17A1 which emits fluorescence corresponding to the amount of the first analyte 9A, a second which emits fluorescence corresponding to the amount of second analyte 9B indicator 17B1 If, consisting of.

For example, a first indicator 17A to be filled in the concave portion 23A comprises a phosphor 81A for generating a fluorescence of a first wavelength corresponding to the amount of glucose, which is the first analyte 9A, a second filling in the concave portion 23B indicator 17B of, including phosphor 81B that emits fluorescence of a second wavelength corresponding to the hydrogen ion concentration of the second analyte 9B. Phosphor 81A includes a phenyl boronic acid derivative, phosphor 81B includes, for example, CdSe / Zn nanoparticles. When phosphors 81A and phosphor 81B are both receiving the excitation light 375 nm, it emits fluorescence in response to the glucose concentration or the hydrogen ion concentration of the analyte.

The calculation unit 2C, in response to a second hydrogen ion concentration indicator 17B was measured (pH), etc., a concentration of glucose first indicator 17A is measured may be corrected. Even if the first indicator 17A and the second indicator 17B contain the same phosphor at different concentrations, it is possible to density correction of the measured analyte.

Depending on the analyte to be measured, the type of indicator 17 is selected. Further, the fluorescence sensor with three or more recesses may have different indicators on the respective recesses. The indicator, for example, phosphor when measuring potassium ions, including crown ether derivative having a fluorescent residue (fluorescence wavelength: 500 nm) can be used, when measuring oxygen, tris (4 , 7-diphenyl-1,10-phenanthroline) perchlorate ruthenium (II) phosphor comprising molecules (fluorescence wavelength: 530 nm) can be used.

Fluorosensor 4E multi-sensing type has the effect of such a fluorescent sensor 4, further, it is possible to measure a plurality of analytes in a single sensor, it is possible to accurately measure a more environmental or biological information. Since it is possible to further measured analyte concentration correction it is more accurate.

Type of measurement to the analyte, i.e. the type of indicator may be changing the wavelength of the excitation light to be irradiated. That is, the first indicator 17A is irradiated with excitation light E1 of the first wavelength, the second indicator 17B may be excitation light E2 of the second wavelength is irradiated. For example, phenylboronic acid derivative is irradiated with first excitation light E1 having a wavelength of 375nm to the first indicator 17A including a phosphor, the second indicator 17B containing GFP a (green fluorescent protein) derivative as phosphor a second excitation light E2 having a wavelength of 490nm may be irradiated.

To irradiated with excitation light of different wavelengths, the sensor portion 10E of the fluorescent sensor. 4E, the first is a light emitting element LED15A1 for generating excitation light E1 of the first wavelength, the excitation light of the second wavelength E2 the second only to have a LED15B1 a light emitting element for generating.

Or it may be used a wavelength conversion filter 80 as the sensor portion 10F of the fluorescence sensor 4F of Modification 3 shown in FIG. 12. For example, the first excitation light E1 having a wavelength of wavelength 375nm which LED15L occurs, the first indicator 17A, as it is irradiated, the wavelength converting filter 80 for excitation light E1 is the second indicator 17B which LED15L generated converted to 460nm excitation light E2 is a second wavelength illuminating the.

Further, the indicator 17A as the sensor portion 10G of the fluorescence sensor 4G of Modification 4 shown in FIG. 13, 17B is a first phosphor that generates first fluorescence F1 corresponding to the amount of the first analyte 9A including 81A and the second phosphor 81B for generating a second fluorescent F2 corresponding to the amount of second analyte 9B, it may be the same.

In this case, the first PD element 12A in which the first phosphor 81A is covered with the first filter 14A1 for selectively transmitting the first fluorescent F1 generated, the second phosphor 81B occurs a second PD element 12B covered with a second filter 14B1 for selectively transmitting a second fluorescent F2 which, if it has a, it is possible to multi-sensing.

The present invention is not limited to the embodiments or the like described above, within the range not changing the gist of the present invention, various changes and modifications are possible.

This application is intended to application Japanese Patent Application No. 2012-103605, filed in Japanese on April 27, 2012 as a basis for claiming priority, the above disclosure, the present specification, claims, and those cited in the drawings.

Claims (8)

  1. A substrate portion where the photoelectric conversion element is formed to output a plurality of recesses are formed, the detection signal received fluorescence to the wall surface of the respective recesses on the main surface,
    Covering the photoelectric conversion element, and a filter that blocks excitation light,
    Is disposed within each of said recesses, when receiving the excitation light and the indicator for generating the fluorescence corresponding to the analyte amount,
    Covering the opening of the recess, but to block the external light entering into the indicator, the analyte and light-shielding layer to pass,
    Fluorescence sensor, characterized by comprising a light emitting element for irradiating excitation light to the indicator from the bottom side of the recess.
  2. Fluorescence sensor according to claim 1, characterized in that the bottom surface of the plurality of recesses, each said light emitting elements are arranged.
  3. The substrate portion, a plurality of light emitting elements and a first substrate portion disposed on the main surface, the first substrate portion and the plurality which is the main surface and the joint through holes second being formed fluorescence sensor according to claim 2 and the substrate portion, characterized in that consist.
  4. A plurality of recesses through hole of the substrate portion, wherein the light emitting element is bonded to the substrate portion so as to cover the region immediately below the opposite side of the opening and opening said covered by the light shielding layer of the plurality of through-holes fluorescent sensor according to claim 1, characterized in that are.
  5. Wherein the plurality of indicators, the first indicator includes a first phosphor that generates first fluorescence corresponding to the amount of the first analyte, a second fluorescence corresponding to the amount of second analyte fluorescence sensor according to any one of claims 1 to 4, and a second indicator includes a second phosphor, characterized in that it consists of generating a.
  6. Wherein the first indicator excitation light of a first wavelength is irradiated, fluorescence sensor according to claim 5, the excitation light of the second wavelength, characterized in that it is irradiated to the second indicator.
  7. The indicator comprises a second phosphor that generates the second fluorescence corresponding to the amount of the first phosphor and the second analyte that generates a first fluorescence corresponding to the amount of the first analyte It includes,
    Second transmitted through the first photoelectric conversion elements covered with a first filter that transmits the first fluorescence wherein said first phosphor is generated, the second fluorescence which said second phosphor is generated fluorescence sensor according to any one of claims 1 to 4 for the second photoelectric conversion elements covered with filter, characterized in that it has a.
  8. Said substrate portion, the filter, the indicator comprises a light shielding layer and the light emitting element needle tip having a sensor portion including a connector portion for fitting portion and fitting of the main body portion arranged outside the body, the , fluorescence sensor according to claim 1, which is a needle-type sensor for measuring the body analyte.
PCT/JP2013/062368 2012-04-27 2013-04-26 Fluorescence sensor WO2013161991A1 (en)

Priority Applications (2)

Application Number Priority Date Filing Date Title
JP2012-103605 2012-04-27
JP2012103605 2012-04-27

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Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH11500825A (en) * 1995-02-21 1999-01-19 アーサー・イー・コルヴィン・ジュニアー Optical fluorescence sensor
US20040161853A1 (en) * 2003-02-13 2004-08-19 Zhongping Yang Implantable chemical sensor with rugged optical coupler
WO2010119916A1 (en) * 2009-04-13 2010-10-21 Olympus Corporation Fluorescence sensor, needle-type fluorescence sensor, and method for measuring analyte

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH11500825A (en) * 1995-02-21 1999-01-19 アーサー・イー・コルヴィン・ジュニアー Optical fluorescence sensor
US20040161853A1 (en) * 2003-02-13 2004-08-19 Zhongping Yang Implantable chemical sensor with rugged optical coupler
WO2010119916A1 (en) * 2009-04-13 2010-10-21 Olympus Corporation Fluorescence sensor, needle-type fluorescence sensor, and method for measuring analyte

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