WO2013161991A1 - Fluorescence sensor - Google Patents

Fluorescence sensor Download PDF

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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
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WO
WIPO (PCT)
Prior art keywords
fluorescence
indicator
sensor
analyte
light
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
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Filing date
Publication date
Application filed by オリンパス株式会社, テルモ株式会社 filed Critical オリンパス株式会社
Publication of WO2013161991A1 publication Critical patent/WO2013161991A1/en

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
    • A61B5/00—Measuring for diagnostic purposes; Identification of persons
    • A61B5/145—Measuring characteristics of blood in vivo, e.g. gas concentration, pH value; Measuring characteristics of body fluids or tissues, e.g. interstitial fluid, cerebral tissue
    • A61B5/14503—Measuring 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
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
    • A61B5/00—Measuring for diagnostic purposes; Identification of persons
    • A61B5/0059—Measuring for diagnostic purposes; Identification of persons using light, e.g. diagnosis by transillumination, diascopy, fluorescence
    • A61B5/0071—Measuring for diagnostic purposes; Identification of persons using light, e.g. diagnosis by transillumination, diascopy, fluorescence by measuring fluorescence emission
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
    • A61B5/00—Measuring for diagnostic purposes; Identification of persons
    • A61B5/145—Measuring characteristics of blood in vivo, e.g. gas concentration, pH value; Measuring characteristics of body fluids or tissues, e.g. interstitial fluid, cerebral tissue
    • A61B5/14532—Measuring 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
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64—Fluorescence; Phosphorescence
    • G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64—Fluorescence; Phosphorescence
    • G01N21/645—Specially adapted constructive features of fluorimeters
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
    • A61B2562/00—Details of sensors; Constructional details of sensor housings or probes; Accessories for sensors
    • A61B2562/12—Manufacturing methods specially adapted for producing sensors for in-vivo measurements
    • 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
    • G01N2021/6417—Spectrofluorimetric devices
    • G01N2021/6421—Measuring at two or more wavelengths
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64—Fluorescence; Phosphorescence
    • G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
    • G01N2021/6439—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" with indicators, stains, dyes, tags, labels, marks
    • G01N2021/6441—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" with indicators, stains, dyes, tags, labels, marks with two or more labels
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
    • G01N21/77—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator
    • G01N2021/7769—Measurement method of reaction-produced change in sensor
    • G01N2021/7786—Fluorescence
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
    • G01N21/77—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator
    • G01N21/78—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator producing a change of colour
    • G01N21/80—Indicating pH value

Definitions

  • the present invention relates to a fluorescent sensor that measures the concentration of an analyte, and more particularly to a fluorescent sensor that is a micro-fluorescence spectrophotometer manufactured using semiconductor manufacturing technology and MEMS technology.
  • Various analyzers have been developed to confirm the presence of analytes in liquids, that is, substances to be measured, or to measure concentrations.
  • a fluorescent dye whose properties change due to the presence of the analyte and generates fluorescence and a solution to be measured containing the analyte are injected, and excitation light is irradiated to increase the fluorescence intensity from the fluorescent dye.
  • Fluorescence spectrophotometers that measure analyte concentration by measuring are known.
  • a small fluorescent spectrophotometer has a photodetector and an indicator containing a fluorescent dye. And, by irradiating the indicator into which the analyte in the solution to be measured can enter the excitation light from the light source, the fluorescent dye in the indicator generates a fluorescence having a light amount corresponding to the analyte concentration in the solution to be measured, The fluorescence is received by the photodetector.
  • the photodetector is a photoelectric conversion element and outputs an electrical signal corresponding to the amount of received light. The analyte concentration in the solution to be measured is measured from this electrical signal.
  • microfluorescence spectrophotometer manufactured using semiconductor manufacturing technology and MEMS technology.
  • the microfluorometer is referred to as a “fluorescence sensor”.
  • a fluorescent sensor 110 shown in FIGS. 1 and 2 is disclosed in US Pat. No. 5,039,490.
  • the fluorescence sensor 110 includes an optical plate-like portion 105 having a transparent support substrate 101 that can transmit excitation light, a photoelectric conversion element 103 that converts fluorescence into an electrical signal, and a condensing function portion 105A that collects excitation light.
  • the indicator 117 is configured to generate fluorescence by interacting with the analyte 9 by the excitation light, and the cover layer 109.
  • the photoelectric conversion element 103 is formed on a substrate 103A made of, for example, silicon.
  • the substrate 103A does not transmit excitation light.
  • the fluorescence sensor 110 has a void region 120 that can transmit excitation light around the photoelectric conversion element 103.
  • Fluorescence F is generated by the interaction between the condensed excitation light E and the analyte 9 that has entered the indicator 117. Part of the generated fluorescence enters the photoelectric conversion element 103, and a signal such as a current or voltage proportional to the fluorescence intensity, that is, the concentration of the analyte 9, is generated in the photoelectric conversion element 103.
  • the excitation light does not enter the photoelectric conversion element 103 due to the action of a filter (not shown) formed on the photoelectric conversion element 103.
  • the photodiode which is the photoelectric conversion element 103 is formed on the substrate 103A laminated on the transparent support substrate 101, and the optical plate portion 105 and the indicator 117 are formed thereon. Are stacked.
  • the above-described fluorescent sensor 110 has the gap region 120 which is a passage of excitation light and the region of the photoelectric conversion element 103 on the same plane. For this reason, if the area of the gap region 120, which is a passage, is increased in order to guide more excitation light to the indicator 117, the area of the photoelectric conversion element 103 is reduced, and thus the sensitivity of the fluorescent sensor is not increased. . On the other hand, if the area of the photoelectric conversion element 103 is increased in order to increase the detection sensitivity of the photoelectric conversion element 103, the area of the gap region 120, which is the path of the excitation light, is reduced, and the excitation light guided to the indicator 117 is reduced. Therefore, the sensitivity of the fluorescence sensor is not increased. That is, with the fluorescent sensor 110, it is not easy to obtain high detection sensitivity.
  • An object of the present invention is to provide a fluorescent sensor with high detection sensitivity.
  • the fluorescent sensor of one embodiment of the present invention includes a substrate portion on which a plurality of recesses are formed on a main surface, and a photoelectric conversion element that receives fluorescence and outputs a detection signal is formed on a wall surface of each recess, A filter that covers the photoelectric conversion element and blocks excitation light; an indicator that is disposed inside each of the recesses; and that generates the fluorescence corresponding to the amount of analyte when receiving the excitation light; and The light-shielding layer that covers the opening and blocks external light from entering the indicator but through which the analyte passes and a light-emitting element that irradiates the indicator with excitation light from the bottom surface side of the recess.
  • the fluorescence sensor 4 constitutes a sensor system 1 together with the main body 2 and the receiver 3. That is, the sensor system 1 includes a fluorescent sensor 4, a main body 2, and a receiver 3 that receives and stores a signal from the main body 2. Transmission / reception of signals between the main body 2 and the receiver 3 is performed wirelessly or by wire.
  • the needle-type fluorescent sensor 4 includes a needle portion 7 having a needle tip portion 5 having a sensor portion 10 as a main functional portion and an elongated needle body portion 6, and a connector portion integrated with a rear end portion of the needle body portion 6. 8 and. Needle tip 5, needle body 6 and connector 8 may be integrally formed of the same material.
  • the connector part 8 is detachably fitted to the fitting part 2A of the main body part 2.
  • the plurality of wirings 60 extending from the sensor unit 10 of the fluorescent sensor 4 are electrically connected to the main body unit 2 when the connector unit 8 is mechanically fitted to the fitting unit 2A of the main body unit 2. .
  • the main body unit 2 includes a control unit 2B including a CPU that drives and controls the sensor unit 10, and a calculation unit 2C including a CPU that processes a signal output from the sensor unit 10.
  • a control unit 2B including a CPU that drives and controls the sensor unit 10 and a calculation unit 2C including a CPU that processes a signal output from the sensor unit 10.
  • the control unit 2B and the calculation unit 2C may be disposed in the connector unit 8 of the fluorescent sensor 4 or may be disposed in the receiver 3. Further, the control unit 2B and the calculation unit 2C may be the same CPU.
  • the main body 2 when performing wired transmission / reception with the receiver 3, the main body 2 has a signal line instead of a wireless antenna. Further, when the main body 2 has a memory unit having a necessary capacity, the receiver 3 is not necessary.
  • the fluorescence sensor 4 is a needle-type sensor that measures an analyte in the body, and is a short-term subcutaneous indwelling type with a continuous use period of about one week.
  • the collected bodily fluid or the bodily fluid that circulates inside the body via a flow path outside the body may be contacted outside the body without inserting the fluorescent sensor 4 into the body.
  • the sensor unit 10 which is the main functional unit of the fluorescence sensor 4 includes a substrate unit 20 and a light emitting diode (Light Emitting Diode: hereinafter referred to as “LED”) that generates excitation light. 15A and 15B, indicators 17A and 17B that generate fluorescence corresponding to the excitation light and the amount of analyte, and a light shielding layer 18.
  • LED Light Emitting Diode
  • 15A and 15B indicators 17A and 17B that generate fluorescence corresponding to the excitation light and the amount of analyte
  • a light shielding layer 18 In the fluorescent sensor 4, the analyte 9 passes through the light shielding layer 18 and enters the indicator 17 when the light shielding layer 18 comes into contact with blood or body fluid in the living body.
  • the substrate unit 20 includes a wiring substrate unit 30 that is a first substrate unit and a frame-shaped substrate unit that is a second substrate unit in which two through holes 46A and 46B are formed on the main surface 20SA (main surface 20SB). 40 are bonded to each other through an adhesive layer 13.
  • substrate part 20 is a multicavity with two recessed part 23A, 23B in main surface 20SA. That is, the surface of the wiring board portion 30 is the bottom surface of the recesses 23A and 23B, and the wall surfaces of the through holes 46A and 46B of the frame-like substrate portion 40 are the wall surfaces of the recesses 23A and 23B.
  • an insulating layer is appropriately formed on the surface of the substrate portion 20 made of, for example, an N-type semiconductor, that is, the wiring substrate portion 30 and the frame-like substrate portion 40, but is not shown. Further, hereinafter, when referring to each component having the same function, the last letter (A or B) is omitted.
  • each of the through holes 46A and 46B is referred to as a through hole 46.
  • the indicator 17A and the LED 15A are disposed inside the recess 23A, and the indicator 17B and the LED 15B are disposed inside the recess 23B.
  • the light emitting element is not limited to the LED. However, a substantially rectangular chip-shaped LED is preferable from the viewpoints of light generation efficiency, wide wavelength selectivity of excitation light, and generation of only light having a wavelength other than ultraviolet light serving as excitation light.
  • the indicator 17 generates fluorescence with a light amount corresponding to the amount of the analyte 9 by the interaction with the analyte 9 that has entered and the excitation light.
  • the indicator 17 generates fluorescence having a longer wavelength, for example, a wavelength of 460 nm with respect to excitation light having a wavelength of 375 nm.
  • the thickness of the indicator 17 is set to about several tens ⁇ m to 200 ⁇ m.
  • the indicator 17 is composed of a base material containing a fluorescent dye that generates fluorescence having an intensity corresponding to the amount of the analyte 9, that is, the concentration of the analyte in the sample.
  • the fluorescent dye is selected according to the type of the analyte 9 and can be used for any fluorescent dye in which the amount of fluorescence generated according to the amount of the analyte 9 changes reversibly. That is, the fluorescent sensor 4 corresponds to various uses such as an oxygen sensor, a glucose sensor, a pH sensor, an immunosensor, or a microorganism sensor, depending on the selection of the fluorescent dye.
  • the indicator 17 includes, for example, a hydrogel that easily contains water as a base material and contains or is bonded to the fluorescent dye in the hydrogel.
  • Hydrogel components are prepared from acrylic hydrogels prepared by polymerizing polysaccharides such as methylcellulose or dextran, monomers such as (meth) acrylamide, methylolacrylamide, or hydroxyethyl acrylate, or from polyethylene glycol and diisocyanate. Urethane hydrogel can be used.
  • the indicator 17 may be joined to the LED 15 through a transparent protective layer that protects the LED 15.
  • a transparent protective layer silicon oxide, epoxy resin, silicone resin, or transparent amorphous fluororesin can be used.
  • the transparent protective layer is selected from materials having characteristics such as electrical insulation, moisture barrier properties, and good transmittance for excitation light and fluorescence. As a characteristic of the transparent protective layer, it is important that the generation of fluorescence in the layer is small even when irradiated with excitation light. Needless to say, the characteristic that the fluorescence is small is an important characteristic of all the transparent materials of the fluorescent sensor 4 except the indicator.
  • the light shielding layer 18 is a layer formed on the upper surface side of the indicator 17 and having a thickness of several tens of ⁇ m or less.
  • the light shielding layer 18 may be divided for the indicator 17A and the indicator 17B.
  • a photodiode (Photo Diode: hereinafter referred to as “PD”) that receives fluorescence and outputs a detection signal.
  • the element 12A is also formed, and the PD element 12B is also formed on the wall surface of the through hole 46B. That is, the light receiving portion 12 ⁇ / b> D of the PD element 12 is provided so as to surround the indicator 17, and is formed so that the light receiving surface faces the indicator 17.
  • the light receiving unit 12D is referred to as a PD element 12.
  • the PD element 12 may be formed on the entire wall surface, but may be formed only in a region facing the indicator 17 in order to efficiently receive only fluorescence.
  • the PD element 12 may be formed on all four wall surfaces, or may be formed on only a part of the surfaces.
  • the PD element 12 may be formed on at least a part of the wall surface of the recess 23.
  • the photoelectric conversion element may be a photoconductor (photoconductor), a phototransistor (Phototransistor, PT), or the like.
  • Filters 14A and 14B are disposed so as to cover the PD elements 12A and 12B formed on the wall surface.
  • the filter 14 covering at least a part of the PD elements 12A and 12B is a high-pass absorption filter that blocks excitation light but allows fluorescence having a wavelength longer than that of the excitation light to pass.
  • a silicon layer or a silicon carbide layer is suitable as a material for such a filter.
  • the transmittance of the silicon layer and the silicon carbide layer is 10 ⁇ 5 % or less at the excitation light wavelength of 375 nm, whereas the transmittance is 10% or more at the fluorescence wavelength of 460 nm (the transmittance of the excitation light wavelength).
  • the filter 14 may be a bandpass filter having a high fluorescence wavelength transmittance.
  • the filter 14 may be arrange
  • the wiring board portion 30 is a wiring layer comprising three wirings 51A, 52, 51B for transmitting detection signals from the PD elements 12A, 12B, and wirings 53, 54 for transmitting drive signals to the LEDs 15A, 15B. 50.
  • the wiring 52 is connected by partially introducing an N-type impurity such as phosphorus or arsenic into the surface of the frame-shaped substrate portion 40, which is an N-type semiconductor, to form a low-resistance region 12H with higher conductivity. Yes.
  • each wiring is arrange
  • the wiring 51 and the wiring 51 ⁇ / b> B may be connected to any one of the plurality of wirings 60 that transmit signals to the main body 2.
  • the material of the frame-shaped substrate portion 40 is preferably single crystal silicon in order to form the PD element 12 on the frame-shaped substrate portion 40, but may be glass or ceramic.
  • a semiconductor thin film such as polysilicon or amorphous silicon of several microns or less is formed on the surface of the recess 23 and the surface of one main surface 20SB of the frame-shaped substrate portion 40, and the PD element 12 is formed thereon.
  • the excitation light generated by the LED 15 is applied to the fluorescent dye in the indicator 17.
  • a part of the fluorescence generated by the interaction of the fluorescent dye with the analyte 9 passes through the filter 14 and reaches the PD element 12 and is converted into a detection signal.
  • the fluorescence sensor 4 has a high detection sensitivity because the PD element 12 formed on the wall surface surrounding the indicator 17 detects fluorescence.
  • the fluorescence sensor 4 Furthermore, in the fluorescence sensor 4, PD elements 12A and 12B are formed on the wall surfaces of the two recesses 23A and 23B, respectively. For this reason, the wall surface area, that is, the area of the light receiving portion 12D of the PD element 12 is large. Further, since the distance from the center of the recess 23 to the light receiving portion of the PD element 12 is short, the fluorescence generated by the fluorescent dye reaches the PD element 12 without being attenuated. For this reason, the fluorescence sensor 4 has a high signal output and a high dynamic sensitivity with a good S / N ratio, and therefore has high detection sensitivity.
  • 6A to 6E are partial cross-sectional views of the region of one fluorescent sensor 4, but in the actual process, a large number of sensor portions 10 are collectively formed as a wafer process.
  • the conductive (N-type) silicon wafer 40W is etched through the mask layer 71 to obtain a large number of through holes 46A, 46B is formed.
  • Various known methods can be used for etching.
  • the size of the opening of the through hole 46 is designed according to the specifications. However, since the location is the needle tip portion 5, the plurality of openings have an elongated shape such as 150 ⁇ m in length and 500 ⁇ m in width, for example.
  • the one opening is preferably about 10 ⁇ m to several hundred ⁇ m.
  • the wall surface of the through hole 46 is perpendicular to the main surface, but the wall surface may have a predetermined angle, that is, a tapered shape, as will be described later.
  • the tapered recess can be produced, for example, by wet etching.
  • the PD element 12 is formed on the wall surface of the through hole 46. That is, the ion implantation process is performed from four directions in a state where the silicon wafer 40W on which the mask layer 72 is formed is inclined by 5 degrees to 30 degrees.
  • the conditions for implanting boron (B) are acceleration voltage: 10 to 100 keV and implantation amount: about 1 ⁇ 10 15 cm ⁇ 2 .
  • the filter 14 is formed by the CVD method on the PD element 12 on the wall surface of the through hole 46 of the silicon wafer 40W.
  • a conductive (N-type) silicon wafer 30 ⁇ / b> W to be the wiring board unit 30 is prepared.
  • an insulating film such as an oxide film is formed on the silicon wafer 30W, drive signals are sent to the wirings 51, 52, 51B and the LEDs 15A, 15B for transmitting the detection signals from the PD element 12.
  • Wirings 53 and 54 for supply are formed by sputtering or vapor deposition.
  • the silicon wafer 40W is turned upside down, and the main surface 20SB of the silicon wafer 40W is bonded to the silicon wafer 30W through the adhesive layer 13.
  • the through hole 46 of the frame-shaped substrate portion 40 becomes a recess 23 having a bottom surface.
  • LEDs 15A and 15B are disposed on the bottom surfaces of the recesses 23A and 23B of the bonded wafer 20W.
  • a bonding method using an optically transparent acrylic resin or silicone resin, or various bonding methods such as a flip chip bonding method can be used.
  • the indicator 17A, 17B is arrange
  • the light shielding layer 18 is formed on the indicators 17A and 17B, and the sensor substrate 10W is completed.
  • the light shielding layer 18 has a submicron pore structure, an inorganic thin film such as metal or ceramic, or a composite structure with hydrogels in which carbon black is mixed in a base material of organic polymer such as polyimide or polyurethane, or Further, a resin obtained by mixing carbon black into an analyte-permeable polymer such as celluloses or polyacrylamide, or a resin obtained by laminating them is used.
  • a large number of sensor units 10 are manufactured at once by separating the sensor substrate 10W into individual pieces. And the fluorescence sensor 4 is completed by joining the sensor part 10 with the front-end
  • the manufacturing method of the fluorescent sensor is not limited to this, and the LED 15 and the like are arranged in the recess 23 after joining the separated wiring board part 30 and the separated frame-like board part 40.
  • a method such as setting may be used.
  • the first silicon wafer may be processed so that the extended portion of the wiring board portion 30 constitutes the needle main body portion 6 of the needle portion 7, or the separately prepared needle main body portion 6 and the fluorescence sensor 4 are provided. You may comprise the needle part 7 by joining the needle front-end
  • the fluorescent sensor 4 can be mass-produced by a wafer process. For this reason, the fluorescence sensor 4 can provide stable quality at low cost.
  • the LED 15 emits pulsed excitation light having a center wavelength of around 375 nm at intervals of, for example, once every 30 seconds.
  • the pulse current to the LED 15 is 1 mA to 100 mA
  • the pulse width of light emission is 101 ms to 100 ms.
  • the excitation light generated by the LED 15 enters the indicator 17.
  • the indicator 17 emits fluorescence having an intensity corresponding to the amount of the analyte 9.
  • the analyte 9 passes through the light shielding layer 18 and enters and exits the indicator 17.
  • a part of the fluorescence generated by the indicator 17 enters the PD element 12 through the filter 14. Fluorescence is photoelectrically converted in the PD element 12 to generate photogenerated charges, which are output as detection signals. Part of the excitation light generated by the LED 15 is incident on the wall surface of the recess 23, but hardly enters the PD element 12 due to the action of the filter 14.
  • the calculation unit 2C of the main body 2 performs calculation processing based on the detection signal, that is, the current caused by the photogenerated charge from the PD element 12 or the voltage caused by the accumulated photogenerated charge. Calculate the amount of light.
  • the fluorescence sensor 4 has two sets of sensor function units capable of independently measuring the analyte, that is, two sets of the LED 15 and the PD element 12. For this reason, the fluorescence sensor 4 can exhibit a specific effect by various driving methods.
  • the LED 15A and the LED 15B may be caused to emit light alternately.
  • the signal output decreases, the fluorescent sensor 4 can perform measurement for a longer time (longer life).
  • the measurement can be continued if the other is operating normally, and thus the fluorescence sensor 4 is improved in reliability. Further, usually, measurement is performed using one sensor function unit, and the other may be used for backup.
  • the indicator 17A and the indicator 17B may be in different deterioration states by changing the integrated light amount irradiated to the LED 15A and the integrated light amount irradiated to the LED 15B. Since it is possible to correct a decrease in the amount of fluorescent light due to deterioration, the fluorescent sensor 4 becomes more accurate.
  • the fluorescent sensor 4 having two sets of sensor function units has effects such as high output, high reliability, long life, and high accuracy by selecting a control method according to the purpose.
  • the fluorescence sensor 4A of the second embodiment will be described. Since the fluorescence sensor 4A is similar to the fluorescence sensor 4 of the first embodiment, the same components are denoted by the same reference numerals and description thereof is omitted.
  • the sensor unit 10A includes a frame-shaped substrate unit 40A and a wiring substrate unit 30A on which one LED 15L is mounted to join the substrate unit 20A. It is composed. That is, one LED 15L irradiates excitation light from below to the two indicators 17A and 17B filled in the two recesses 23A and 23B.
  • the PD element 12A and the PD element 12B are connected, and a detection signal is output via the common wirings 51 and 52.
  • the fluorescent sensor 4A has a simple structure because one LED 15L is used as a common excitation light source for the two indicators 17A and 17B.
  • the fluorescence sensor 4B of the third embodiment will be described. Since the fluorescence sensor 4B is similar to the fluorescence sensors 4 and 4A, the same components are denoted by the same reference numerals and description thereof is omitted.
  • the substrate portion 20 ⁇ / b> B of the sensor portion 10 ⁇ / b> B is a region directly below the opening on the side opposite to the opening covered with the light shielding layer 18.
  • One LED 15L is joined to the frame-shaped substrate portion 40B so as to cover the frame.
  • the LED 15L covers the region immediately below the opening on the main surface 20SB side of the two through holes 46A and 46B.
  • the bottom surfaces of the through holes 46A and 46B are constituted by the upper surface of the LED 15L.
  • the bottom surfaces of the through holes 46A and 46B may be the transparent adhesive layer 13.
  • the wiring layer 50 which comprises the wiring connected with PD element 12A, 12B and LED15L is arrange
  • the light leakage prevention layer 19 disposed so as to cover the bottom surface (lower surface) and the side surface of the LED 15L is excited by the excitation light emitted from the bottom surface and the side surface and the excitation reflected by the second main surface 20SB of the frame-shaped substrate portion 40B. Prevent light from leaking outside. That is, the light leakage prevention layer 19 has a function similar to that of the light shielding layer 18, but does not require analyte permeability.
  • the fluorescence sensor 4B has the effects of the fluorescence sensors 4 and 4A, and further has a simple configuration, so that it is easier to manufacture.
  • the substrate portion 20C constituting the sensor portion 10C is integrally made of silicon which is a semiconductor. That is, the recesses 23A and 23B of the substrate portion 20C are recesses formed by an etching method.
  • etching method a wet etching method using a tetramethylammonium hydroxide (TMAH) aqueous solution, a potassium hydroxide (KOH) aqueous solution, or the like is preferable, but dry etching such as reactive ion etching (RIE) or chemical dry etching (CDE) is used.
  • TMAH tetramethylammonium hydroxide
  • KOH potassium hydroxide
  • CDE chemical dry etching
  • PD elements 12A and 12B are formed on the wall surfaces of the recesses 23A and 23B, respectively.
  • the concave portion 23 having the wall surface taper not only has a larger area for forming the PD element than the concave portion having a vertical wall surface, but also facilitates the formation of the PD element 12.
  • the PD elements 12A and 13A and the LEDs 15A and 15B are connected to the wiring layer 50 of the second main surface 20SB through the through wiring.
  • the fluorescence sensor 4C has the effects of the fluorescence sensor 4 and is simpler in configuration, and therefore easier to manufacture.
  • the sensor units 10 to 10C have the two concave portions 23A and 23B that are rectangular in a plan view (cross-sectional shape).
  • the number, that is, the number of indicators 17 may be three or more.
  • it may have four recesses 23A to 23D like a fluorescent sensor 4D (sensor unit 10D) of the modification shown in FIG. 10B.
  • the cross-sectional shape of the plurality of recesses 23 may be a polygon such as a hexagon or an ellipse, or various shapes such as a combination thereof. But you can.
  • the fluorescence sensor has high detection sensitivity.
  • the intensity of the excitation light generated by the LED 15 has an in-plane distribution.
  • the fluorescence intensity emitted from the indicator 17 in the region irradiated with the high-intensity excitation light is high.
  • the fluorescence intensity emitted by the indicator 17 in the region where the intensity of the incident excitation light is low is low. That is, if the intensity of the excitation light incident on the indicator 17 has a large in-plane distribution, the detection sensitivity of the fluorescent sensor may be reduced, or deterioration with time may be accelerated.
  • the intensity distribution of the excitation light incident on the indicator 17 can be averaged by the opening area, shape, arrangement, and the like of the plurality of recesses 23.
  • Fluorescence sensors 4D to 4D2 of Modification 1 can not only have higher detection sensitivity, but also suppress the deterioration of indicator 17 in addition to the effects of fluorescence sensors 4 to 4C.
  • the plurality of indicators 17 and the plurality of PD elements 12 measure the same type of analyte 9, for example, glucose. That is, the fluorescence sensors 4 to 4D2 correspond to various applications such as an oxygen sensor, a glucose sensor, a pH sensor, an immunosensor, or a microorganism sensor, depending on the selection of the fluorescent dye. However, only one type of analyte 9 can be measured.
  • the sensor unit 10E of the fluorescence sensor 4E of the modification 2 shown in FIG. 11 is a multi-sensing type capable of measuring a plurality of types of analytes.
  • the indicator generates a first indicator 17A1 that generates fluorescence according to the amount of the first analyte 9A, and a second indicator 17B1 that generates fluorescence according to the amount of the second analyte 9B. And consist of
  • the first indicator 17A that fills the recess 23A includes the phosphor 81A that generates fluorescence of the first wavelength according to the amount of glucose that is the first analyte 9A, and the second indicator that fills the recess 23B.
  • the indicator 17B includes a phosphor 81B that generates fluorescence of the second wavelength corresponding to the hydrogen ion concentration, which is the second analyte 9B.
  • the phosphor 81A includes a phenylboronic acid derivative, and the phosphor 81B includes, for example, CdSe / Zn nanoparticles.
  • both the phosphor 81A and the phosphor 81B receive the excitation light of 375 nm, the phosphor 81A and the phosphor 81B generate fluorescence corresponding to the glucose concentration or the hydrogen ion concentration as the analyte.
  • the computing unit 2C may correct the glucose concentration measured by the first indicator 17A according to the hydrogen ion concentration (pH) measured by the second indicator 17B. Even when the first indicator 17A and the second indicator 17B contain the same phosphor at different concentrations, the concentration of the measured analyte can be corrected.
  • the type of indicator 17 is selected according to the analyte to be measured. Further, a fluorescent sensor having three or more recesses may have a different indicator in each recess.
  • the indicator for example, when measuring potassium ion, a phosphor (fluorescence wavelength: 500 nm) including a crown ether derivative having a fluorescent residue can be used, and when measuring oxygen, Tris (4 , 7-diphenyl-1,10-phenanthroline) a phosphor containing a ruthenium (II) perchlorate molecule (fluorescence wavelength: 530 nm) can be used.
  • the multi-sensing type fluorescence sensor 4E has the effect of the fluorescence sensor 4 and the like, and moreover, since a plurality of analytes can be measured with one sensor, the environment and biological information can be measured more accurately. Furthermore, since the measured analyte concentration can be corrected, the accuracy is higher.
  • the wavelength of the excitation light to be irradiated may be changed depending on the type of analyte to be measured, that is, the type of indicator. That is, the first indicator 17A may be irradiated with the excitation light E1 having the first wavelength, and the second indicator 17B may be irradiated with the excitation light E2 having the second wavelength.
  • the first indicator 17A including a phenylboronic acid derivative as a phosphor is irradiated with a first excitation light E1 having a wavelength of 375 nm
  • the second indicator 17B including a GFP (green fluorescent protein) derivative as a phosphor You may irradiate the 2nd excitation light E2 of wavelength 490nm.
  • the sensor unit 10E of the fluorescence sensor 4E includes an LED 15A1 that is a first light emitting element that generates excitation light E1 of the first wavelength, and excitation light E2 of the second wavelength.
  • LED15B1 which is the 2nd light emitting element which generate
  • the wavelength conversion filter 80 may be used as in the sensor unit 10F of the fluorescence sensor 4F of Modification 3 shown in FIG.
  • the wavelength conversion filter 80 emits the excitation light E1 of the first wavelength of 375 nm generated by the LED 15L to the first indicator 17A as it is, and the excitation light E1 generated by the LED 15L to the second indicator 17B. Is converted into excitation light E2 having a second wavelength of 460 nm and irradiated.
  • the indicators 17A and 17B generate the first phosphor F1 that generates the first fluorescence F1 according to the amount of the first analyte 9A.
  • produces the 2nd fluorescence F2 according to the quantity of the 2nd analyte 9B may be sufficient.
  • the first PD element 12A covered by the first filter 14A1 that selectively transmits the first fluorescence F1 generated by the first phosphor 81A and the second phosphor 81B are generated. If it has 2nd PD element 12B covered with 2nd filter 14B1 which permeate

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

蛍光センサFluorescent sensor
　本発明は、アナライトの濃度を計測する蛍光センサに関し、特に半導体製造技術およびＭＥＭＳ技術を用いて作製される微小蛍光分光光度計である蛍光センサに関する。 The present invention relates to a fluorescent sensor that measures the concentration of an analyte, and more particularly to a fluorescent sensor that is a micro-fluorescence spectrophotometer manufactured using semiconductor manufacturing technology and MEMS technology.
　液体中のアナライトすなわち被計測物質の存在確認、または、濃度を測定するための様々な分析装置が開発されている。例えば、一定容量の透明容器に、アナライトの存在によって性質が変化し蛍光を発生する蛍光色素と、アナライトを含む被計測溶液とを注入し、励起光を照射し蛍光色素からの蛍光強度を計測することによりアナライト濃度を計測する蛍光分光光度計が知られている。 Various analyzers have been developed to confirm the presence of analytes in liquids, that is, substances to be measured, or to measure concentrations. For example, in a transparent container with a certain volume, a fluorescent dye whose properties change due to the presence of the analyte and generates fluorescence and a solution to be measured containing the analyte are injected, and excitation light is irradiated to increase the fluorescence intensity from the fluorescent dye. Fluorescence spectrophotometers that measure analyte concentration by measuring are known.
　小型の蛍光分光光度計は、光検出器と蛍光色素を含有したインジケータとを有している。そして、被計測溶液中のアナライトが進入可能なインジケータに光源からの励起光を照射することで、インジケータ内の蛍光色素が被計測溶液中のアナライト濃度に応じた光量の蛍光を発生し、その蛍光を光検出器が受光する。光検出器は光電変換素子であり、受光した光量に応じた電気信号を出力する。この電気信号から被計測溶液中のアナライト濃度が測定される。 A small fluorescent spectrophotometer has a photodetector and an indicator containing a fluorescent dye. And, by irradiating the indicator into which the analyte in the solution to be measured can enter the excitation light from the light source, the fluorescent dye in the indicator generates a fluorescence having a light amount corresponding to the analyte concentration in the solution to be measured, The fluorescence is received by the photodetector. The photodetector is a photoelectric conversion element and outputs an electrical signal corresponding to the amount of received light. The analyte concentration in the solution to be measured is measured from this electrical signal.
　近年、微量試料中のアナライトを計測するために、半導体製造技術およびＭＥＭＳ技術を用いて作製される微小蛍光分光光度計が提案されている。以下、微小蛍光光度計のことを、「蛍光センサ」と呼ぶ。 In recent years, in order to measure an analyte in a small amount of sample, a micro-fluorescence spectrophotometer manufactured using semiconductor manufacturing technology and MEMS technology has been proposed. Hereinafter, the microfluorometer is referred to as a “fluorescence sensor”.
　例えば、図１および図２に示す蛍光センサ１１０が米国特許第５０３９４９０号明細書に開示されている。蛍光センサ１１０は、励起光が透過可能な透明支持基板１０１と、蛍光を電気信号に変換する光電変換素子１０３と、励起光を集光する集光機能部１０５Ａとを有する光学板状部１０５と、アナライト９と相互作用することによって励起光の入射により蛍光を発生するインジケータ１１７と、カバー層１０９と、から構成されている。 For example, a fluorescent sensor 110 shown in FIGS. 1 and 2 is disclosed in US Pat. No. 5,039,490. The fluorescence sensor 110 includes an optical plate-like portion 105 having a transparent support substrate 101 that can transmit excitation light, a photoelectric conversion element 103 that converts fluorescence into an electrical signal, and a condensing function portion 105A that collects excitation light. The indicator 117 is configured to generate fluorescence by interacting with the analyte 9 by the excitation light, and the cover layer 109.
　光電変換素子１０３は、例えばシリコンからなる基板１０３Ａに形成されている。基板１０３Ａは励起光を透過しない。このため、蛍光センサ１１０では、光電変換素子１０３の周囲に励起光が透過可能な空隙領域１２０を有している。 The photoelectric conversion element 103 is formed on a substrate 103A made of, for example, silicon. The substrate 103A does not transmit excitation light. For this reason, the fluorescence sensor 110 has a void region 120 that can transmit excitation light around the photoelectric conversion element 103.
　すなわち、空隙領域１２０を透過した励起光Ｅだけが、集光機能部１０５Ａの作用により、インジケータ１１７中の、光電変換素子１０３の上部付近に集光される。集光された励起光Ｅと、インジケータ１１７の内部に進入したアナライト９の相互作用により、蛍光Ｆが発生する。発生した蛍光の一部は光電変換素子１０３に入射し、光電変換素子１０３において蛍光強度、つまりアナライト９の濃度に比例した電流または電圧などの信号が発生する。なお励起光は、光電変換素子１０３上に形成されたフィルタ（不図示）の作用により、光電変換素子１０３には入射しない。 That is, only the excitation light E transmitted through the gap region 120 is condensed near the upper portion of the photoelectric conversion element 103 in the indicator 117 by the action of the condensing function unit 105A. Fluorescence F is generated by the interaction between the condensed excitation light E and the analyte 9 that has entered the indicator 117. Part of the generated fluorescence enters the photoelectric conversion element 103, and a signal such as a current or voltage proportional to the fluorescence intensity, that is, the concentration of the analyte 9, is generated in the photoelectric conversion element 103. The excitation light does not enter the photoelectric conversion element 103 due to the action of a filter (not shown) formed on the photoelectric conversion element 103.
　以上の説明のように、蛍光センサ１１０は、透明支持基板１０１の上に積層される基板１０３Ａに光電変換素子１０３であるフォトダイオードを形成し、その上に、光学板状部１０５およびインジケータ１１７が積層されている。 As described above, in the fluorescence sensor 110, the photodiode which is the photoelectric conversion element 103 is formed on the substrate 103A laminated on the transparent support substrate 101, and the optical plate portion 105 and the indicator 117 are formed thereon. Are stacked.
　しかし、上記の蛍光センサ１１０は、励起光の通路である空隙領域１２０と光電変換素子１０３の領域とを、同一平面上に有する。このため、より多くの励起光をインジケータ１１７に導光するために通路である空隙領域１２０の面積を広くすると、光電変換素子１０３の面積が狭くなるため、蛍光センサの感度を高めることにはならない。反対に光電変換素子１０３の検出感度を高くするために光電変換素子１０３の面積を広くすると、励起光の通路である空隙領域１２０の面積が狭くなりインジケータ１１７に導光する励起光が減少してしまうために、やはり蛍光センサの感度を高めることにはならない。すなわち、蛍光センサ１１０では、高い検出感度を得ることは容易ではなかった。 However, the above-described fluorescent sensor 110 has the gap region 120 which is a passage of excitation light and the region of the photoelectric conversion element 103 on the same plane. For this reason, if the area of the gap region 120, which is a passage, is increased in order to guide more excitation light to the indicator 117, the area of the photoelectric conversion element 103 is reduced, and thus the sensitivity of the fluorescent sensor is not increased. . On the other hand, if the area of the photoelectric conversion element 103 is increased in order to increase the detection sensitivity of the photoelectric conversion element 103, the area of the gap region 120, which is the path of the excitation light, is reduced, and the excitation light guided to the indicator 117 is reduced. Therefore, the sensitivity of the fluorescence sensor is not increased. That is, with the fluorescent sensor 110, it is not easy to obtain high detection sensitivity.
　本発明は検出感度の高い蛍光センサを提供することを目的とする。 An object of the present invention is to provide a fluorescent sensor with high detection sensitivity.
　本発明の一態様の蛍光センサは、主面に複数の凹部が形成されており、それぞれの凹部の壁面に蛍光を受光し検出信号を出力する光電変換素子が形成されている基板部と、前記光電変換素子を覆う、励起光を遮断するフィルタと、それぞれの前記凹部の内部に配設されている、前記励起光を受光するとアナライト量に応じた前記蛍光を発生するインジケータと、前記凹部の開口を覆う、前記インジケータへの外光進入を遮断するが、アナライトは通過する遮光層と、前記凹部の底面側から前記インジケータに励起光を照射する発光素子と、を具備する。 The fluorescent sensor of one embodiment of the present invention includes a substrate portion on which a plurality of recesses are formed on a main surface, and a photoelectric conversion element that receives fluorescence and outputs a detection signal is formed on a wall surface of each recess, A filter that covers the photoelectric conversion element and blocks excitation light; an indicator that is disposed inside each of the recesses; and that generates the fluorescence corresponding to the amount of analyte when receiving the excitation light; and The light-shielding layer that covers the opening and blocks external light from entering the indicator but through which the analyte passes and a light-emitting element that irradiates the indicator with excitation light from the bottom surface side of the recess.
従来の蛍光センサの断面図である。It is sectional drawing of the conventional fluorescence sensor. 従来の蛍光センサの構造を説明するための分解図である。It is an exploded view for demonstrating the structure of the conventional fluorescence sensor. 第１実施形態の蛍光センサを有するセンサシステムを説明するための説明図である。It is explanatory drawing for demonstrating the sensor system which has the fluorescence sensor of 1st Embodiment. 第１実施形態の蛍光センサの構造を説明するための分解図である。It is an exploded view for demonstrating the structure of the fluorescence sensor of 1st Embodiment. 第１実施形態の蛍光センサの断面図である。It is sectional drawing of the fluorescence sensor of 1st Embodiment. 第１実施形態の蛍光センサの製造方法を説明するための断面図である。It is sectional drawing for demonstrating the manufacturing method of the fluorescence sensor of 1st Embodiment. 第１実施形態の蛍光センサの製造方法を説明するための断面図である。It is sectional drawing for demonstrating the manufacturing method of the fluorescence sensor of 1st Embodiment. 第１実施形態の蛍光センサの製造方法を説明するための断面図である。It is sectional drawing for demonstrating the manufacturing method of the fluorescence sensor of 1st Embodiment. 第１実施形態の蛍光センサの製造方法を説明するための断面図である。It is sectional drawing for demonstrating the manufacturing method of the fluorescence sensor of 1st Embodiment. 第１実施形態の蛍光センサの製造方法を説明するための断面図である。It is sectional drawing for demonstrating the manufacturing method of the fluorescence sensor of 1st Embodiment. 第２実施形態の蛍光センサの構造を説明するための分解断面図である。It is a disassembled sectional view for demonstrating the structure of the fluorescence sensor of 2nd Embodiment. 第３実施形態の蛍光センサの構造を説明するための分解断面図である。It is a disassembled sectional view for demonstrating the structure of the fluorescence sensor of 3rd Embodiment. 第４実施形態の蛍光センサの製造方法を説明するための断面図である。It is sectional drawing for demonstrating the manufacturing method of the fluorescence sensor of 4th Embodiment. 変形例１の蛍光センサのインジケータの配置を説明するための上面図である。It is a top view for demonstrating arrangement | positioning of the indicator of the fluorescence sensor of the modification 1. 変形例１の蛍光センサのインジケータの配置を説明するための上面図である。It is a top view for demonstrating arrangement | positioning of the indicator of the fluorescence sensor of the modification 1. 変形例１の蛍光センサのインジケータの配置を説明するための上面図である。It is a top view for demonstrating arrangement | positioning of the indicator of the fluorescence sensor of the modification 1. 変形例１の蛍光センサのインジケータの配置を説明するための上面図である。It is a top view for demonstrating arrangement | positioning of the indicator of the fluorescence sensor of the modification 1. 変形例２の蛍光センサを説明するための断面図である。It is sectional drawing for demonstrating the fluorescence sensor of the modification 2. 変形例３の蛍光センサを説明するための断面図である。It is sectional drawing for demonstrating the fluorescence sensor of the modification 3. 変形例４の蛍光センサを説明するための断面図である。It is sectional drawing for demonstrating the fluorescence sensor of the modification 4.
＜第１実施形態＞
　以下、本発明の第１実施形態の蛍光センサ４について説明する。 <First Embodiment>
Hereinafter, the fluorescence sensor 4 according to the first embodiment of the present invention will be described.
　図３に示すように、蛍光センサ４は、本体部２およびレシーバー３と、ともにセンサシステム１を構成する。すなわち、センサシステム１は、蛍光センサ４と、本体部２と、本体部２からの信号を受信し記憶するレシーバー３と、を有する。本体部２とレシーバー３との間の信号の送受信は無線または有線で行われる。 As shown in FIG. 3, the fluorescence sensor 4 constitutes a sensor system 1 together with the main body 2 and the receiver 3. That is, the sensor system 1 includes a fluorescent sensor 4, a main body 2, and a receiver 3 that receives and stores a signal from the main body 2. Transmission / reception of signals between the main body 2 and the receiver 3 is performed wirelessly or by wire.
　針型の蛍光センサ４は、主要機能部であるセンサ部１０を有する針先端部５と細長い針本体部６とを有する針部７と、針本体部６の後端部と一体化したコネクタ部８と、を具備する。針先端部５、針本体部６、およびコネクタ部８は同一材料により一体形成されていてもよい。 The needle-type fluorescent sensor 4 includes a needle portion 7 having a needle tip portion 5 having a sensor portion 10 as a main functional portion and an elongated needle body portion 6, and a connector portion integrated with a rear end portion of the needle body portion 6. 8 and. Needle tip 5, needle body 6 and connector 8 may be integrally formed of the same material.
　コネクタ部８は、本体部２の嵌合部２Ａと着脱自在に嵌合する。蛍光センサ４のセンサ部１０から延設された複数の配線６０は、コネクタ部８が本体部２の嵌合部２Ａと機械的に嵌合することにより、本体部２と電気的に接続される。 The connector part 8 is detachably fitted to the fitting part 2A of the main body part 2. The plurality of wirings 60 extending from the sensor unit 10 of the fluorescent sensor 4 are electrically connected to the main body unit 2 when the connector unit 8 is mechanically fitted to the fitting unit 2A of the main body unit 2. .
　本体部２は、センサ部１０の駆動および制御などを行うＣＰＵ等からなる制御部２Ｂと、センサ部１０から出力された信号を処理するＣＰＵ等からなる演算部２Ｃと、を有する。なお、制御部２Ｂまたは演算部２Ｃの少なくともいずれかが、蛍光センサ４のコネクタ部８等に配設されていてもよいし、レシーバー３に配設されていてもよい。また、制御部２Ｂ及び演算部２Ｃが同一のＣＰＵであってもよい。 The main body unit 2 includes a control unit 2B including a CPU that drives and controls the sensor unit 10, and a calculation unit 2C including a CPU that processes a signal output from the sensor unit 10. Note that at least one of the control unit 2B and the calculation unit 2C may be disposed in the connector unit 8 of the fluorescent sensor 4 or may be disposed in the receiver 3. Further, the control unit 2B and the calculation unit 2C may be the same CPU.
　なお、レシーバー３との間を有線送受信する場合には、本体部２は無線アンテナに代えて信号線を有する。また、本体部２が必要な容量のメモリ部を有する場合にはレシーバー３は不要である。 In addition, when performing wired transmission / reception with the receiver 3, the main body 2 has a signal line instead of a wireless antenna. Further, when the main body 2 has a memory unit having a necessary capacity, the receiver 3 is not necessary.
　蛍光センサ４は本体部２と嵌合した状態で、被検者自身が体表面から穿刺して針先端部５が体内に留置される。そして、例えば体液中のグルコース濃度を連続して測定し、レシーバー３のメモリに記憶する。すなわち、蛍光センサ４は、体内のアナライトを計測する針型センサであり、連続使用期間が一週間程度の短期皮下留置型である。しかし、蛍光センサ４を体内に挿入しないで、採取した体液、または体外の流路を介して体内と循環する体液を、体外において接触させてもよい。 When the fluorescent sensor 4 is fitted to the main body 2, the subject himself punctures the body surface and the needle tip 5 is left in the body. For example, the glucose concentration in the body fluid is continuously measured and stored in the memory of the receiver 3. That is, the fluorescence sensor 4 is a needle-type sensor that measures an analyte in the body, and is a short-term subcutaneous indwelling type with a continuous use period of about one week. However, the collected bodily fluid or the bodily fluid that circulates inside the body via a flow path outside the body may be contacted outside the body without inserting the fluorescent sensor 4 into the body.
　そして、図４、図５に示すように、蛍光センサ４の主機能部であるセンサ部１０は、基板部２０と、励起光を発生する発光素子である発光ダイオード（Light Emitting Diode：以下「ＬＥＤ」ともいう）１５Ａ、１５Ｂと、励起光とアナライト量とに応じた蛍光を発生するインジケータ１７Ａ、１７Ｂと、遮光層１８と、を有する。蛍光センサ４では、遮光層１８が生体中の血液または体液と接触することにより、アナライト９が遮光層１８を通過してインジケータ１７に進入する。 As shown in FIGS. 4 and 5, the sensor unit 10 which is the main functional unit of the fluorescence sensor 4 includes a substrate unit 20 and a light emitting diode (Light Emitting Diode: hereinafter referred to as “LED”) that generates excitation light. 15A and 15B, indicators 17A and 17B that generate fluorescence corresponding to the excitation light and the amount of analyte, and a light shielding layer 18. In the fluorescent sensor 4, the analyte 9 passes through the light shielding layer 18 and enters the indicator 17 when the light shielding layer 18 comes into contact with blood or body fluid in the living body.
　基板部２０は、第１の基板部である配線基板部３０と、主面２０ＳＡ（主面２０ＳＢ）に２個の貫通孔４６Ａ、４６Ｂが形成された第２の基板部である枠状基板部４０と、を接着層１３を介して接合することにより作製されている。このため、基板部２０は、主面２０ＳＡに２つの凹部２３Ａ、２３Ｂがあるマルチキャビティである。すなわち、配線基板部３０の表面が、凹部２３Ａ、２３Ｂの底面であり、枠状基板部４０の貫通孔４６Ａ、４６Ｂの壁面が、凹部２３Ａ、２３Ｂの壁面である。 The substrate unit 20 includes a wiring substrate unit 30 that is a first substrate unit and a frame-shaped substrate unit that is a second substrate unit in which two through holes 46A and 46B are formed on the main surface 20SA (main surface 20SB). 40 are bonded to each other through an adhesive layer 13. For this reason, the board | substrate part 20 is a multicavity with two recessed part 23A, 23B in main surface 20SA. That is, the surface of the wiring board portion 30 is the bottom surface of the recesses 23A and 23B, and the wall surfaces of the through holes 46A and 46B of the frame-like substrate portion 40 are the wall surfaces of the recesses 23A and 23B.
　なお、例えばＮ型半導体からなる基板部２０、すなわち配線基板部３０および枠状基板部４０の表面等には適宜、絶縁層が形成されるが図示していない。また、以下、同じ機能の構成要素のそれぞれをいうときは末尾の一文字（ＡまたはＢ）を省略する。例えば、貫通孔４６Ａ、４６Ｂのそれぞれを貫通孔４６という。 For example, an insulating layer is appropriately formed on the surface of the substrate portion 20 made of, for example, an N-type semiconductor, that is, the wiring substrate portion 30 and the frame-like substrate portion 40, but is not shown. Further, hereinafter, when referring to each component having the same function, the last letter (A or B) is omitted. For example, each of the through holes 46A and 46B is referred to as a through hole 46.
　凹部２３Ａの内部には、インジケータ１７ＡとＬＥＤ１５Ａとが配設されており、凹部２３Ｂの内部にはインジケータ１７ＢとＬＥＤ１５Ｂとが配設されている。発光素子としては、ＬＥＤに限られるものではない。しかし、光発生効率、励起光の波長選択性の広さ、および励起光となる紫外線以外の波長の光を僅かしか発生しないことなどの観点から、略矩形のチップ状のＬＥＤが好ましい。 The indicator 17A and the LED 15A are disposed inside the recess 23A, and the indicator 17B and the LED 15B are disposed inside the recess 23B. The light emitting element is not limited to the LED. However, a substantially rectangular chip-shaped LED is preferable from the viewpoints of light generation efficiency, wide wavelength selectivity of excitation light, and generation of only light having a wavelength other than ultraviolet light serving as excitation light.
　インジケータ１７は、進入してきたアナライト９との相互作用および励起光によりアナライト９の量に応じた光量の蛍光を発生する。例えば、インジケータ１７は波長３７５ｎｍの励起光に対して、より長波長の例えば波長４６０ｎｍの蛍光を発生する。インジケータ１７の厚さは数十μｍ～２００μｍ程度に設定されている。インジケータ１７は、アナライト９の量、すなわち試料中のアナライト濃度に応じた強度の蛍光を発生する蛍光色素が含まれたベース材料から構成されている。 The indicator 17 generates fluorescence with a light amount corresponding to the amount of the analyte 9 by the interaction with the analyte 9 that has entered and the excitation light. For example, the indicator 17 generates fluorescence having a longer wavelength, for example, a wavelength of 460 nm with respect to excitation light having a wavelength of 375 nm. The thickness of the indicator 17 is set to about several tens μm to 200 μm. The indicator 17 is composed of a base material containing a fluorescent dye that generates fluorescence having an intensity corresponding to the amount of the analyte 9, that is, the concentration of the analyte in the sample.
　蛍光色素は、アナライト９の種類に応じて選択され、アナライト９の量に応じて発生する蛍光の光量が可逆的に変化する蛍光色素ならば、どのようなものにも使用できる。すなわち、蛍光センサ４は、蛍光色素の選択によって、酸素センサ、グルコースセンサ、ｐＨセンサ、免疫センサ、または微生物センサなど、多様な用途に対応している。 The fluorescent dye is selected according to the type of the analyte 9 and can be used for any fluorescent dye in which the amount of fluorescence generated according to the amount of the analyte 9 changes reversibly. That is, the fluorescent sensor 4 corresponds to various uses such as an oxygen sensor, a glucose sensor, a pH sensor, an immunosensor, or a microorganism sensor, depending on the selection of the fluorescent dye.
　インジケータ１７は、例えば、含水し易いハイドロゲルをベース材料として、ハイドロゲル内に上記蛍光色素を含有するまたは結合されている。ハイドロゲルの成分としてはメチルセルロースもしくはデキストランなどの多糖類、（メタ）アクリルアミド、メチロールアクリルアミド、もしくはヒドルキシエチルアクリレート等のモノマーを重合して作製するアクリル系ハイドロゲル、またはポリエチレングリコールとジイソシアネートから作製するウレタン系ハイドロゲルなどを用いることができる。 The indicator 17 includes, for example, a hydrogel that easily contains water as a base material and contains or is bonded to the fluorescent dye in the hydrogel. Hydrogel components are prepared from acrylic hydrogels prepared by polymerizing polysaccharides such as methylcellulose or dextran, monomers such as (meth) acrylamide, methylolacrylamide, or hydroxyethyl acrylate, or from polyethylene glycol and diisocyanate. Urethane hydrogel can be used.
　インジケータ１７はＬＥＤ１５を保護する透明な保護層を介してＬＥＤ１５と接合されていてもよい。透明保護層としては、酸化シリコン、エポキシ樹脂、シリコーン樹脂、または透明な非晶性フッ素樹脂などが使用可能である。透明保護層は、電気的絶縁性を有すること、水分遮断性を有すること、励起光および蛍光に対して良好な透過率を有すること、などの特性を有する材料から選択される。透明保護層の特性として、励起光が照射されても層中での蛍光の発生が小さいことが重要である。なお、この蛍光が小さいという特性は、インジケータを除いた蛍光センサ４の全ての透明材料の重要特性であることは言うまでもない。 The indicator 17 may be joined to the LED 15 through a transparent protective layer that protects the LED 15. As the transparent protective layer, silicon oxide, epoxy resin, silicone resin, or transparent amorphous fluororesin can be used. The transparent protective layer is selected from materials having characteristics such as electrical insulation, moisture barrier properties, and good transmittance for excitation light and fluorescence. As a characteristic of the transparent protective layer, it is important that the generation of fluorescence in the layer is small even when irradiated with excitation light. Needless to say, the characteristic that the fluorescence is small is an important characteristic of all the transparent materials of the fluorescent sensor 4 except the indicator.
　遮光層１８は、インジケータ１７の上部表面側に形成された、厚さが数十μｍ以下の層である。なお、遮光層１８はインジケータ１７Ａ用とインジケータ１７Ｂ用とで分割されていてもよい。 The light shielding layer 18 is a layer formed on the upper surface side of the indicator 17 and having a thickness of several tens of μm or less. The light shielding layer 18 may be divided for the indicator 17A and the indicator 17B.
　一方、枠状基板部４０の貫通孔４６Ａの壁面、すなわち基板部２０の凹部２３Ａの壁面には、蛍光を受光し検出信号を出力する光電変換素子であるフォトダイオード（Photo Diode：以下「ＰＤ」ともいう）素子１２Ａが形成されており、同様に貫通孔４６Ｂの壁面にもＰＤ素子１２Ｂが形成されている。すなわち、ＰＤ素子１２の受光部１２Ｄは、インジケータ１７を囲むように設けられ、受光面がインジケータ１７に向くように形成されている。以下、受光部１２ＤをＰＤ素子１２という。 On the other hand, on the wall surface of the through hole 46A of the frame-shaped substrate portion 40, that is, the wall surface of the recess portion 23A of the substrate portion 20, a photodiode (Photo Diode: hereinafter referred to as “PD”) that receives fluorescence and outputs a detection signal. The element 12A is also formed, and the PD element 12B is also formed on the wall surface of the through hole 46B. That is, the light receiving portion 12 </ b> D of the PD element 12 is provided so as to surround the indicator 17, and is formed so that the light receiving surface faces the indicator 17. Hereinafter, the light receiving unit 12D is referred to as a PD element 12.
　ＰＤ素子１２は、壁面の全体に形成されていてもよいが、蛍光のみを効率的に受光するために、インジケータ１７との対向領域にのみに形成してもよい。またＰＤ素子１２は、４面ある壁面の全てに形成されていてもよいし、一部の面のみに形成されていてもよい。 The PD element 12 may be formed on the entire wall surface, but may be formed only in a region facing the indicator 17 in order to efficiently receive only fluorescence. The PD element 12 may be formed on all four wall surfaces, or may be formed on only a part of the surfaces.
　すなわち、ＰＤ素子１２は、凹部２３の壁面の少なくとも一部に形成されていればよい。また、光電変換素子としては、フォトコンダクタ（光導電体）、またはフォトトランジスタ（Photo Transistor、PT）などでもよい。 That is, the PD element 12 may be formed on at least a part of the wall surface of the recess 23. The photoelectric conversion element may be a photoconductor (photoconductor), a phototransistor (Phototransistor, PT), or the like.
　壁面に形成された、それぞれのＰＤ素子１２Ａ、１２Ｂを覆うようにフィルタ１４Ａ、１４Ｂが配設されている。ＰＤ素子１２Ａ、１２Ｂの少なくとも一部を覆うフィルタ１４は励起光を遮断するが、励起光よりも長波長の蛍光は通すハイパス型の吸収型フィルタである。このようなフィルタの材料としては、シリコン層または炭化シリコン層が好適である。例えば、シリコン層および炭化シリコン層は、励起光波長の３７５ｎｍでは透過率は１０－５％以下であるのに対して、蛍光波長の４６０ｎｍでは透過率１０％以上と、（励起光波長の透過率／蛍光波長の透過率）の比として６桁以上の透過率選択性を有する。なお、フィルタ１４として蛍光波長の透過率が高いバンドパスフィルタであってもよい。また、ＰＤ素子１２を保護する透明層、例えば酸化シリコン層を介してフィルタ１４が配設されていてもよい。 Filters 14A and 14B are disposed so as to cover the PD elements 12A and 12B formed on the wall surface. The filter 14 covering at least a part of the PD elements 12A and 12B is a high-pass absorption filter that blocks excitation light but allows fluorescence having a wavelength longer than that of the excitation light to pass. As a material for such a filter, a silicon layer or a silicon carbide layer is suitable. For example, the transmittance of the silicon layer and the silicon carbide layer is 10 −5 % or less at the excitation light wavelength of 375 nm, whereas the transmittance is 10% or more at the fluorescence wavelength of 460 nm (the transmittance of the excitation light wavelength). (Transmittance of fluorescence wavelength) and the transmittance selectivity of 6 digits or more. Note that the filter 14 may be a bandpass filter having a high fluorescence wavelength transmittance. Moreover, the filter 14 may be arrange | positioned through the transparent layer which protects PD element 12, for example, a silicon oxide layer.
　配線基板部３０は、ＰＤ素子１２Ａ、１２Ｂからの検出信号を伝達するための３本の配線５１Ａ、５２、５１Ｂ、およびＬＥＤ１５Ａ、１５Ｂに駆動信号を伝達するための配線５３、５４からなる配線層５０を有する。配線５２は、Ｎ型半導体である枠状基板部４０の表面に部分的にＮ型不純物、例えばリンまたはヒ素等を導入して、より導電率の高い低抵抗領域１２Ｈを形成し、接続されている。なお、ＬＥＤ１５Ａ、１５Ｂを別々に駆動する場合には、それぞれの配線が配設される。また、配線５１と配線５１Ｂとは接続されてから本体部２に信号を伝達する複数の配線６０のいずれかと接続されてもよい。 The wiring board portion 30 is a wiring layer comprising three wirings 51A, 52, 51B for transmitting detection signals from the PD elements 12A, 12B, and wirings 53, 54 for transmitting drive signals to the LEDs 15A, 15B. 50. The wiring 52 is connected by partially introducing an N-type impurity such as phosphorus or arsenic into the surface of the frame-shaped substrate portion 40, which is an N-type semiconductor, to form a low-resistance region 12H with higher conductivity. Yes. In addition, when driving LED15A, 15B separately, each wiring is arrange | positioned. In addition, the wiring 51 and the wiring 51 </ b> B may be connected to any one of the plurality of wirings 60 that transmit signals to the main body 2.
　枠状基板部４０の材料は、ＰＤ素子１２を枠状基板部４０に形成するためには単結晶シリコンが好ましいが、ガラスまたはセラミック等でもよい。その場合、凹部２３の表面および枠状基板部４０の主面のひとつ２０ＳＢの表面に、数ミクロン以下のポリシリコンまたはアモルファスシリコン等の半導体薄膜が形成され、それにＰＤ素子１２が形成される。 The material of the frame-shaped substrate portion 40 is preferably single crystal silicon in order to form the PD element 12 on the frame-shaped substrate portion 40, but may be glass or ceramic. In that case, a semiconductor thin film such as polysilicon or amorphous silicon of several microns or less is formed on the surface of the recess 23 and the surface of one main surface 20SB of the frame-shaped substrate portion 40, and the PD element 12 is formed thereon.
　ＬＥＤ１５が発生した励起光は、インジケータ１７中の蛍光色素に照射される。そして、蛍光色素がアナライト９との相互作用により発生した蛍光の一部は、フィルタ１４を通過してＰＤ素子１２に到達し、検出信号に変換される。 The excitation light generated by the LED 15 is applied to the fluorescent dye in the indicator 17. A part of the fluorescence generated by the interaction of the fluorescent dye with the analyte 9 passes through the filter 14 and reaches the PD element 12 and is converted into a detection signal.
　蛍光センサ４は、インジケータ１７を取り囲む壁面に形成されたＰＤ素子１２により蛍光を検出するために、検出感度が高い。 The fluorescence sensor 4 has a high detection sensitivity because the PD element 12 formed on the wall surface surrounding the indicator 17 detects fluorescence.
　さらに、蛍光センサ４では２つの凹部２３Ａ、２３Ｂの壁面にそれぞれＰＤ素子１２Ａ、１２Ｂが形成されている。このため、壁面面積すなわちＰＤ素子１２の受光部１２Ｄの面積が広い。さらに、凹部２３の中心からＰＤ素子１２の受光部までの距離が短いために蛍光色素が発生した蛍光が減衰することなくＰＤ素子１２に到達する。このため、蛍光センサ４は、信号出力が高く、Ｓ／Ｎ比のよい広いダイナミックレンジのため、検出感度が高い。 Furthermore, in the fluorescence sensor 4, PD elements 12A and 12B are formed on the wall surfaces of the two recesses 23A and 23B, respectively. For this reason, the wall surface area, that is, the area of the light receiving portion 12D of the PD element 12 is large. Further, since the distance from the center of the recess 23 to the light receiving portion of the PD element 12 is short, the fluorescence generated by the fluorescent dye reaches the PD element 12 without being attenuated. For this reason, the fluorescence sensor 4 has a high signal output and a high dynamic sensitivity with a good S / N ratio, and therefore has high detection sensitivity.
　次に、蛍光センサ４の製造方法について簡単に説明する。なお、図６Ａ～図６Ｅでは１個の蛍光センサ４の領域の部分断面図であるが、実際の工程では、ウエハプロセスとして一括して多数のセンサ部１０を形成する。 Next, a method for manufacturing the fluorescent sensor 4 will be briefly described. 6A to 6E are partial cross-sectional views of the region of one fluorescent sensor 4, but in the actual process, a large number of sensor portions 10 are collectively formed as a wafer process.
　最初に、図６Ａに示すように、枠状基板部４０の作製では、導電性（Ｎ型）のシリコンウエハ４０Ｗに対して、マスク層７１を介してエッチングが行われ、多数の貫通孔４６Ａ、４６Ｂが形成される。エッチングには公知の各種の方法を用いることができる。 First, as shown in FIG. 6A, in the production of the frame-shaped substrate portion 40, the conductive (N-type) silicon wafer 40W is etched through the mask layer 71 to obtain a large number of through holes 46A, 46B is formed. Various known methods can be used for etching.
　なお、貫通孔４６の開口の大きさは仕様に応じて設計されるが、配設箇所が針先端部５であるために、複数の開口は、例えば、縦１５０μｍ、横５００μｍのように細長い形状の領域に配置されることが好ましく、一つの開口は１０μｍから数百μｍ程度の大きさが好ましい。 The size of the opening of the through hole 46 is designed according to the specifications. However, since the location is the needle tip portion 5, the plurality of openings have an elongated shape such as 150 μm in length and 500 μm in width, for example. The one opening is preferably about 10 μm to several hundred μm.
　また、蛍光センサ４では、貫通孔４６の壁面は主面に垂直であるが、後述するように、壁面は所定の角度、すなわちテーパーのある形状であってもよい。テーパー形状の凹部は例えばウエットエッチングにより作製できる。 Further, in the fluorescent sensor 4, the wall surface of the through hole 46 is perpendicular to the main surface, but the wall surface may have a predetermined angle, that is, a tapered shape, as will be described later. The tapered recess can be produced, for example, by wet etching.
　次に、図６Ｂに示すように、貫通孔４６の壁面にＰＤ素子１２が形成される。すなわち、マスク層７２が形成されたシリコンウエハ４０Ｗを５度～３０度に傾けた状態で、４方向からイオン注入処理が行われる。例えば、ほう素（Ｂ）を注入する場合の条件は、加速電圧：１０～１００ｋｅＶ、注入量：１×１０１５ｃｍ－２程度である。 Next, as shown in FIG. 6B, the PD element 12 is formed on the wall surface of the through hole 46. That is, the ion implantation process is performed from four directions in a state where the silicon wafer 40W on which the mask layer 72 is formed is inclined by 5 degrees to 30 degrees. For example, the conditions for implanting boron (B) are acceleration voltage: 10 to 100 keV and implantation amount: about 1 × 10 15 cm −2 .
　そして、シリコンウエハ４０Ｗの貫通孔４６の壁面のＰＤ素子１２上に、フィルタ１４がＣＶＤ法により、形成される。 Then, the filter 14 is formed by the CVD method on the PD element 12 on the wall surface of the through hole 46 of the silicon wafer 40W.
　別途、配線基板部３０となる導電性（Ｎ型）のシリコンウエハ３０Ｗが準備される。シリコンウエハ３０Ｗには、図示していないが、酸化膜等の絶縁膜が形成された後、ＰＤ素子１２からの検出信号を送信するための配線５１、５２、５１ＢおよびＬＥＤ１５Ａ、１５Ｂに駆動信号を供給するための配線５３、５４が、スパッタ法または蒸着法等により形成される。 Separately, a conductive (N-type) silicon wafer 30 </ b> W to be the wiring board unit 30 is prepared. Although an insulating film such as an oxide film is formed on the silicon wafer 30W, drive signals are sent to the wirings 51, 52, 51B and the LEDs 15A, 15B for transmitting the detection signals from the PD element 12. Wirings 53 and 54 for supply are formed by sputtering or vapor deposition.
　図６Ｃに示すように、シリコンウエハ４０Ｗが上下反転されて、シリコンウエハ４０Ｗの主面２０ＳＢが接着層１３を介してシリコンウエハ３０Ｗと接合される。 As shown in FIG. 6C, the silicon wafer 40W is turned upside down, and the main surface 20SB of the silicon wafer 40W is bonded to the silicon wafer 30W through the adhesive layer 13.
　図６Ｄに示すように、２枚のウエハが接合された接合ウエハ２０Ｗでは、枠状基板部４０の貫通孔４６は底面のある凹部２３となる。次に接合ウエハ２０Ｗの凹部２３Ａ、２３Ｂのそれぞれの底面に、ＬＥＤ１５Ａ、１５Ｂが配設される。ＬＥＤ１５の配設には、光学的に透明なアクリル樹脂もしくはシリコーン樹脂などを使用した接着法、または、フリップチップボンディング法などの各種接合法などの方法が使用可能である。 As shown in FIG. 6D, in the bonded wafer 20W in which two wafers are bonded, the through hole 46 of the frame-shaped substrate portion 40 becomes a recess 23 having a bottom surface. Next, LEDs 15A and 15B are disposed on the bottom surfaces of the recesses 23A and 23B of the bonded wafer 20W. For the arrangement of the LED 15, a bonding method using an optically transparent acrylic resin or silicone resin, or various bonding methods such as a flip chip bonding method can be used.
　そして、図６Ｅに示すように、接合ウエハ２０Ｗの凹部２３Ａ、２３Ｂのそれぞれの内部に、インジケータ１７Ａ、１７Ｂが配設される。 And as shown to FIG. 6E, the indicator 17A, 17B is arrange | positioned inside each of the recessed parts 23A, 23B of the bonded wafer 20W.
　次に、遮光層１８がインジケータ１７Ａ、１７Ｂ上に形成されセンサ基板１０Ｗが完成する。遮光層１８には、サブミクロンサイズのポア構造からなる、金属、セラミック等の無機薄膜または、ポリイミドもしくはポリウレタン等の有機ポリマーの基材にカーボンブラックが混入されたハイドロゲル類とのコンポジット構造、または、セルロース類もしくはポリアクリルアミド等のアナライト透過性ポリマーにカーボンブラックを混入した樹脂、または、それらを積層化した樹脂等を用いる。 Next, the light shielding layer 18 is formed on the indicators 17A and 17B, and the sensor substrate 10W is completed. The light shielding layer 18 has a submicron pore structure, an inorganic thin film such as metal or ceramic, or a composite structure with hydrogels in which carbon black is mixed in a base material of organic polymer such as polyimide or polyurethane, or Further, a resin obtained by mixing carbon black into an analyte-permeable polymer such as celluloses or polyacrylamide, or a resin obtained by laminating them is used.
　センサ基板１０Ｗが個片化されることにより、多数のセンサ部１０が一括して作製される。そして、センサ部１０が、別途作製された、コネクタ部８から延設された針本体部６の先端部と接合されることで、蛍光センサ４が完成する。 A large number of sensor units 10 are manufactured at once by separating the sensor substrate 10W into individual pieces. And the fluorescence sensor 4 is completed by joining the sensor part 10 with the front-end | tip part of the needle | hook main-body part 6 extended from the connector part 8 produced separately.
　蛍光センサの製造方法としては、これに限られるものではなく、個片化された配線基板部３０と、個片化された枠状基板部４０とを接合した後に、凹部２３にＬＥＤ１５等を配設する等の方法を用いてもよい。 The manufacturing method of the fluorescent sensor is not limited to this, and the LED 15 and the like are arranged in the recess 23 after joining the separated wiring board part 30 and the separated frame-like board part 40. A method such as setting may be used.
　また、配線基板部３０の延設部が針部７の針本体部６を構成するように第１のシリコンウエハを加工してもよいし、別途作製した針本体部６と、蛍光センサ４を有する針先端部５とを接合して針部７を構成してもよい。 Further, the first silicon wafer may be processed so that the extended portion of the wiring board portion 30 constitutes the needle main body portion 6 of the needle portion 7, or the separately prepared needle main body portion 6 and the fluorescence sensor 4 are provided. You may comprise the needle part 7 by joining the needle front-end | tip part 5 which has.
　以上の説明のように、蛍光センサ４は、ウエハプロセスにより一括大量生産が可能である。このため、蛍光センサ４は、安価に安定した品質を提供できる。 As described above, the fluorescent sensor 4 can be mass-produced by a wafer process. For this reason, the fluorescence sensor 4 can provide stable quality at low cost.
　次に、蛍光センサ４の動作の一例を説明する。 Next, an example of the operation of the fluorescence sensor 4 will be described.
　ＬＥＤ１５は、例えば３０秒に１回の間隔で中心波長が３７５ｎｍ前後の励起光をパルス発光する。例えば、ＬＥＤ１５へのパルス電流は１ｍＡ～１００ｍＡであり、発光のパルス幅は１０１ｍｓ～１００ｍｓである。 The LED 15 emits pulsed excitation light having a center wavelength of around 375 nm at intervals of, for example, once every 30 seconds. For example, the pulse current to the LED 15 is 1 mA to 100 mA, and the pulse width of light emission is 101 ms to 100 ms.
　ＬＥＤ１５が発生した励起光はインジケータ１７に入射する。インジケータ１７は、アナライト９の量に対応した強度の蛍光を発する。なお、アナライト９は遮光層１８を通過して、インジケータ１７に出入する。 The excitation light generated by the LED 15 enters the indicator 17. The indicator 17 emits fluorescence having an intensity corresponding to the amount of the analyte 9. The analyte 9 passes through the light shielding layer 18 and enters and exits the indicator 17.
　インジケータ１７が発生した蛍光の一部は、フィルタ１４を介して、ＰＤ素子１２に入射する。そして蛍光は、ＰＤ素子１２において光電変換され光発生電荷を生じることで、検出信号として出力される。なお、ＬＥＤ１５が発生する励起光も一部は、凹部２３の壁面に入射するが、フィルタ１４の作用によりＰＤ素子１２へは殆ど入射しない。 A part of the fluorescence generated by the indicator 17 enters the PD element 12 through the filter 14. Fluorescence is photoelectrically converted in the PD element 12 to generate photogenerated charges, which are output as detection signals. Part of the excitation light generated by the LED 15 is incident on the wall surface of the recess 23, but hardly enters the PD element 12 due to the action of the filter 14.
　蛍光センサ４では、本体部２の演算部２Ｃが検出信号、すなわち、ＰＤ素子１２からの光発生電荷に起因する電流または蓄積した光発生電荷に起因する電圧をもとに演算処理を行い、アナライト量を算出する。 In the fluorescence sensor 4, the calculation unit 2C of the main body 2 performs calculation processing based on the detection signal, that is, the current caused by the photogenerated charge from the PD element 12 or the voltage caused by the accumulated photogenerated charge. Calculate the amount of light.
　ここで、蛍光センサ４は、独立してアナライトの測定が可能な２組のセンサ機能部、すなわち、ＬＥＤ１５とＰＤ素子１２のセットを２組、を有する。このため、蛍光センサ４は各種の駆動方法により特有の効果を奏することができる。 Here, the fluorescence sensor 4 has two sets of sensor function units capable of independently measuring the analyte, that is, two sets of the LED 15 and the PD element 12. For this reason, the fluorescence sensor 4 can exhibit a specific effect by various driving methods.
　例えば、励起光の照射によりインジケータの劣化が進行する場合には、ＬＥＤ１５ＡとＬＥＤ１５Ｂとを交互に発光させてもよい。信号出力は低下するが、蛍光センサ４はより長時間の測定（長寿命）が可能となる。 For example, when deterioration of the indicator progresses due to irradiation with excitation light, the LED 15A and the LED 15B may be caused to emit light alternately. Although the signal output decreases, the fluorescent sensor 4 can perform measurement for a longer time (longer life).
　また、一方のセンサ機能部（ＬＥＤ１５またはＰＤ素子１２等）が故障しても他方が正常に動作していれば測定を継続できるため、蛍光センサ４は信頼性が向上する。また、通常は、一方のセンサ機能部を用いて測定を行い、他方をバックアップ用としてもよい。 In addition, even if one sensor function unit (LED 15 or PD element 12 or the like) fails, the measurement can be continued if the other is operating normally, and thus the fluorescence sensor 4 is improved in reliability. Further, usually, measurement is performed using one sensor function unit, and the other may be used for backup.
　さらに、ＬＥＤ１５Ａに照射する積算光量とＬＥＤ１５Ｂに照射する積算光量とを変えることで、インジケータ１７Ａとインジケータ１７Ｂを異なる劣化状態としてもよい。劣化による蛍光光量減少を補正することが可能となるため、蛍光センサ４はより高精度となる。 Further, the indicator 17A and the indicator 17B may be in different deterioration states by changing the integrated light amount irradiated to the LED 15A and the integrated light amount irradiated to the LED 15B. Since it is possible to correct a decrease in the amount of fluorescent light due to deterioration, the fluorescent sensor 4 becomes more accurate.
　以上の説明のように、２組のセンサ機能部を有する蛍光センサ４は、目的に応じて制御方法を選択することにより、高出力、高信頼性、長寿命、高精度等の効果を有する。 As described above, the fluorescent sensor 4 having two sets of sensor function units has effects such as high output, high reliability, long life, and high accuracy by selecting a control method according to the purpose.
＜第２実施形態＞
　次に、第２実施形態の蛍光センサ４Ａについて説明する。蛍光センサ４Ａは第１実施形態の蛍光センサ４と類似しているため同じ構成要素には同じ符号を付し説明は省略する。 <Second Embodiment>
Next, the fluorescence sensor 4A of the second embodiment will be described. Since the fluorescence sensor 4A is similar to the fluorescence sensor 4 of the first embodiment, the same components are denoted by the same reference numerals and description thereof is omitted.
　図７に示すように、第２実施形態の蛍光センサ４Ａでは、センサ部１０Ａは、枠状基板部４０Ａと、１個のＬＥＤ１５Ｌが実装された配線基板部３０Ａとが接合されて基板部２０Ａを構成している。すなわち、２つの凹部２３Ａ、２３Ｂに充填された２つのインジケータ１７Ａ、１７Ｂに、１個のＬＥＤ１５Ｌが下方から励起光を照射する。 As shown in FIG. 7, in the fluorescent sensor 4A of the second embodiment, the sensor unit 10A includes a frame-shaped substrate unit 40A and a wiring substrate unit 30A on which one LED 15L is mounted to join the substrate unit 20A. It is composed. That is, one LED 15L irradiates excitation light from below to the two indicators 17A and 17B filled in the two recesses 23A and 23B.
　なお、図７に示す蛍光センサ４Ａでは、ＰＤ素子１２ＡとＰＤ素子１２Ｂとは接続されており、共通の配線５１、５２を介して検出信号を出力する。 In the fluorescent sensor 4A shown in FIG. 7, the PD element 12A and the PD element 12B are connected, and a detection signal is output via the common wirings 51 and 52.
　蛍光センサ４Ａは、１個のＬＥＤ１５Ｌを２つのインジケータ１７Ａ、１７Ｂの共通の励起光源としているため構造が簡単である。 The fluorescent sensor 4A has a simple structure because one LED 15L is used as a common excitation light source for the two indicators 17A and 17B.
＜第３実施形態＞
　次に、第３実施形態の蛍光センサ４Ｂについて説明する。蛍光センサ４Ｂは蛍光センサ４、４Ａと類似しているため同じ構成要素には同じ符号を付し説明は省略する。 <Third Embodiment>
Next, the fluorescence sensor 4B of the third embodiment will be described. Since the fluorescence sensor 4B is similar to the fluorescence sensors 4 and 4A, the same components are denoted by the same reference numerals and description thereof is omitted.
　図８に示すように、第３実施形態の蛍光センサ４Ｂでは、センサ部１０Ｂの基板部２０Ｂは、貫通孔４６の開口のうち、遮光層１８に覆われた開口と反対側の開口の直下領域を覆うように１個のＬＥＤ１５Ｌが、枠状基板部４０Ｂと接合されて構成されている。 As shown in FIG. 8, in the fluorescent sensor 4 </ b> B of the third embodiment, the substrate portion 20 </ b> B of the sensor portion 10 </ b> B is a region directly below the opening on the side opposite to the opening covered with the light shielding layer 18. One LED 15L is joined to the frame-shaped substrate portion 40B so as to cover the frame.
　すなわち、ＬＥＤ１５Ｌは、２つの貫通孔４６Ａ、４６Ｂの主面２０ＳＢ側の開口の直下領域を覆っている。言い換えれば、貫通孔４６Ａ、４６Ｂの底面はＬＥＤ１５Ｌの上面により構成されている。なお貫通孔４６Ａ、４６Ｂの底面は、透明な接着層１３であってもよい。 That is, the LED 15L covers the region immediately below the opening on the main surface 20SB side of the two through holes 46A and 46B. In other words, the bottom surfaces of the through holes 46A and 46B are constituted by the upper surface of the LED 15L. The bottom surfaces of the through holes 46A and 46B may be the transparent adhesive layer 13.
　そして、ＰＤ素子１２Ａ、１２ＢおよびＬＥＤ１５Ｌと接続された配線を構成する配線層５０が枠状基板部４０Ｂに配設されている。 And the wiring layer 50 which comprises the wiring connected with PD element 12A, 12B and LED15L is arrange | positioned in the frame-shaped board | substrate part 40B.
　ＬＥＤ１５Ｌの底面（下面）および側面を覆うように配設された漏光防止層１９は、底面および側面から出射される励起光、および、枠状基板部４０Ｂの第２の主面２０ＳＢで反射した励起光が外部に漏光するのを防止する。すなわち、漏光防止層１９は遮光層１８と類似した機能を有するが、アナライト透過性は必要ない。 The light leakage prevention layer 19 disposed so as to cover the bottom surface (lower surface) and the side surface of the LED 15L is excited by the excitation light emitted from the bottom surface and the side surface and the excitation reflected by the second main surface 20SB of the frame-shaped substrate portion 40B. Prevent light from leaking outside. That is, the light leakage prevention layer 19 has a function similar to that of the light shielding layer 18, but does not require analyte permeability.
　以上の説明のように、蛍光センサ４Ｂは、蛍光センサ４、４Ａの効果を有し、さらに構成が簡単なため、より製造が容易である。 As described above, the fluorescence sensor 4B has the effects of the fluorescence sensors 4 and 4A, and further has a simple configuration, so that it is easier to manufacture.
＜第４実施形態＞
　次に、第４実施形態の蛍光センサ４Ｃについて説明する。蛍光センサ４Ｃは蛍光センサ４と類似しているため同じ構成要素には同じ符号を付し説明は省略する。 <Fourth embodiment>
Next, the fluorescence sensor 4C of the fourth embodiment will be described. Since the fluorescence sensor 4C is similar to the fluorescence sensor 4, the same components are denoted by the same reference numerals and description thereof is omitted.
　図９に示すように、蛍光センサ４Ｃでは、センサ部１０Ｃを構成する基板部２０Ｃが半導体であるシリコンにより一体的に作製されている。すなわち、基板部２０Ｃの凹部２３Ａ、２３Ｂは、エッチング法により形成された凹部である。 As shown in FIG. 9, in the fluorescence sensor 4C, the substrate portion 20C constituting the sensor portion 10C is integrally made of silicon which is a semiconductor. That is, the recesses 23A and 23B of the substrate portion 20C are recesses formed by an etching method.
　エッチング法としては、水酸化テトラメチルアンモニウム（ＴＭＡＨ）水溶液、水酸化カリウム（ＫＯＨ）水溶液などを用いるウエットエッチング法が望ましいが、反応性イオンエッチング（ＲＩＥ）、ケミカルドライエッチング（ＣＤＥ）などのドライエッチング法も用いることができる
　例えば、シリコンとしてシリコン（１００）面を主面とする基板を用いた場合には、（１１１）面のエッチング速度が（１００）面に比べて遅い異方性エッチングとなるため、の凹部２３Ａ、２３Ｂの壁面は（１１１）面となり、（１００）面との角度θ１は５４．７４度となる。すなわち、壁面はテーパー形状である。 As an etching method, a wet etching method using a tetramethylammonium hydroxide (TMAH) aqueous solution, a potassium hydroxide (KOH) aqueous solution, or the like is preferable, but dry etching such as reactive ion etching (RIE) or chemical dry etching (CDE) is used. For example, when a substrate having a silicon (100) surface as a main surface is used as silicon, anisotropic etching with a slower etching rate of the (111) surface than the (100) surface is performed. Therefore, the wall surfaces of the recesses 23A and 23B are the (111) plane, and the angle θ1 with the (100) plane is 54.74 degrees. That is, the wall surface is tapered.
　そして、凹部２３Ａ、２３Ｂの壁面に、それぞれＰＤ素子１２Ａ、１２Ｂが形成される。壁面テーパーのある凹部２３は、壁面が垂直な凹部に比べてＰＤ素子を形成する面積が広いだけでなく、ＰＤ素子１２の形成が容易である。 PD elements 12A and 12B are formed on the wall surfaces of the recesses 23A and 23B, respectively. The concave portion 23 having the wall surface taper not only has a larger area for forming the PD element than the concave portion having a vertical wall surface, but also facilitates the formation of the PD element 12.
　なお、ＰＤ素子１２Ａ、１３Ａ、ＬＥＤ１５Ａ、１５Ｂは、貫通配線を介して第２の主面２０ＳＢの配線層５０と接続されている。 The PD elements 12A and 13A and the LEDs 15A and 15B are connected to the wiring layer 50 of the second main surface 20SB through the through wiring.
　以上の説明のように、蛍光センサ４Ｃは、蛍光センサ４の効果を有し、さらに構成が簡単なため、より製造が容易である。 As described above, the fluorescence sensor 4C has the effects of the fluorescence sensor 4 and is simpler in configuration, and therefore easier to manufacture.
＜変形例１＞
　図１０Ａに示すように、実施形態１の蛍光センサ４～４Ｃでは、センサ部１０～１０Ｃは２個の平面視（断面形状）が矩形の凹部２３Ａ、２３Ｂを有していたが、凹部２３の数、すなわち、インジケータ１７の数は３個以上であってもよい。例えば、図１０Ｂに示す変形例の蛍光センサ４Ｄ（センサ部１０Ｄ）のように４個の凹部２３Ａ～２３Ｄを有していてもよい。また、図１０Ｃに示す蛍光センサ４Ｄ１（センサ部１０Ｄ１）のように複数の凹部２３の断面形状は６角形等の多角形もしくは楕円形等でもよいし、または、それらを組み合わせたような各種の形状でもよい。また図１０Ｄに示す蛍光センサ４Ｄ２（センサ部１０Ｄ２）のように、形状および大きさが異なる凹部２３を組み合わせてもよい。 <Modification 1>
As shown in FIG. 10A, in the fluorescent sensors 4 to 4C of the first embodiment, the sensor units 10 to 10C have the two concave portions 23A and 23B that are rectangular in a plan view (cross-sectional shape). The number, that is, the number of indicators 17 may be three or more. For example, it may have four recesses 23A to 23D like a fluorescent sensor 4D (sensor unit 10D) of the modification shown in FIG. 10B. Further, like the fluorescent sensor 4D1 (sensor unit 10D1) shown in FIG. 10C, the cross-sectional shape of the plurality of recesses 23 may be a polygon such as a hexagon or an ellipse, or various shapes such as a combination thereof. But you can. Moreover, you may combine the recessed part 23 from which a shape and magnitude | size differ like fluorescence sensor 4D2 (sensor part 10D2) shown to FIG. 10D.
　同一面積のセンサ部であっても凹部２３の数が多いと壁面の面積が増加するため、蛍光センサは検出感度が高くなる、
　ここで、ＬＥＤ１５が発生する励起光の強度には面内分布がある。高強度の励起光が照射される領域のインジケータ１７が発光する蛍光強度は高い。しかし、高強度の励起光が照射されると、インジケータ１７の劣化が促進されるおそれがある。これに対して入射する励起光の強度が低い領域のインジケータ１７が発光する蛍光強度は低い。すなわち、インジケータ１７に入射する励起光の強度に大きな面内分布があると、蛍光センサの検出感度が低下したり、経時劣化が加速したりするおそれがある。 Even if the sensor portions have the same area, if the number of the recesses 23 is large, the area of the wall surface increases. Therefore, the fluorescence sensor has high detection sensitivity.
Here, the intensity of the excitation light generated by the LED 15 has an in-plane distribution. The fluorescence intensity emitted from the indicator 17 in the region irradiated with the high-intensity excitation light is high. However, when high-intensity excitation light is irradiated, deterioration of the indicator 17 may be promoted. On the other hand, the fluorescence intensity emitted by the indicator 17 in the region where the intensity of the incident excitation light is low is low. That is, if the intensity of the excitation light incident on the indicator 17 has a large in-plane distribution, the detection sensitivity of the fluorescent sensor may be reduced, or deterioration with time may be accelerated.
　しかし、蛍光センサ４Ｄ２のように、複数の凹部２３の開口面積、形状および配置等により、インジケータ１７に入射する励起光の強度分布を平均化することもできる。 However, like the fluorescence sensor 4D2, the intensity distribution of the excitation light incident on the indicator 17 can be averaged by the opening area, shape, arrangement, and the like of the plurality of recesses 23.
　変形例１の蛍光センサ４Ｄ～４Ｄ２は、蛍光センサ４～４Ｃが有する効果に加えて、さらに、より検出感度が高いだけでなく、インジケータ１７の劣化進行を抑制することもできる。 Fluorescence sensors 4D to 4D2 of Modification 1 can not only have higher detection sensitivity, but also suppress the deterioration of indicator 17 in addition to the effects of fluorescence sensors 4 to 4C.
＜変形例２～４＞
　すでに説明した蛍光センサ４～４Ｄ２では、複数のインジケータ１７および複数のＰＤ素子１２が同じ種類のアナライト９、例えばグルコースを測定した。すなわち、蛍光センサ４～４Ｄ２は、蛍光色素の選択によって、酸素センサ、グルコースセンサ、ｐＨセンサ、免疫センサ、または微生物センサなど、多様な用途に対応している。しかし、測定できるアナライト９は１種類であった。 <Modifications 2 to 4>
In the fluorescence sensors 4 to 4D2 already described, the plurality of indicators 17 and the plurality of PD elements 12 measure the same type of analyte 9, for example, glucose. That is, the fluorescence sensors 4 to 4D2 correspond to various applications such as an oxygen sensor, a glucose sensor, a pH sensor, an immunosensor, or a microorganism sensor, depending on the selection of the fluorescent dye. However, only one type of analyte 9 can be measured.
　これに対して、図１１に示す変形例２の蛍光センサ４Ｅのセンサ部１０Ｅは複数種類のアナライトを測定可能なマルチセンシング型である。蛍光センサ４Ｅでは、インジケータが、第１のアナライト９Ａの量に応じた蛍光を発生する第１のインジケータ１７Ａ１と、第２のアナライト９Ｂの量に応じた蛍光を発生する第２のインジケータ１７Ｂ１と、からなる。 On the other hand, the sensor unit 10E of the fluorescence sensor 4E of the modification 2 shown in FIG. 11 is a multi-sensing type capable of measuring a plurality of types of analytes. In the fluorescence sensor 4E, the indicator generates a first indicator 17A1 that generates fluorescence according to the amount of the first analyte 9A, and a second indicator 17B1 that generates fluorescence according to the amount of the second analyte 9B. And consist of
　例えば、凹部２３Ａに充填する第１のインジケータ１７Ａが、第１のアナライト９Ａであるグルコースの量に応じた第１の波長の蛍光を発生する蛍光体８１Ａを含み、凹部２３Ｂに充填する第２のインジケータ１７Ｂが、第２のアナライト９Ｂである水素イオン濃度に応じた第２の波長の蛍光を発生する蛍光体８１Ｂを含む。蛍光体８１Aはフェニルボロン酸誘導体を含み、蛍光体８１Ｂは、例えば、ＣｄＳｅ／Ｚｎナノ微粒子を含む。蛍光体８１A及び蛍光体８１Bは共に３７５ｎｍの励起光を受光すると、アナライトであるグルコース濃度または水素イオン濃度に応じた蛍光を発生する。 For example, the first indicator 17A that fills the recess 23A includes the phosphor 81A that generates fluorescence of the first wavelength according to the amount of glucose that is the first analyte 9A, and the second indicator that fills the recess 23B. The indicator 17B includes a phosphor 81B that generates fluorescence of the second wavelength corresponding to the hydrogen ion concentration, which is the second analyte 9B. The phosphor 81A includes a phenylboronic acid derivative, and the phosphor 81B includes, for example, CdSe / Zn nanoparticles. When both the phosphor 81A and the phosphor 81B receive the excitation light of 375 nm, the phosphor 81A and the phosphor 81B generate fluorescence corresponding to the glucose concentration or the hydrogen ion concentration as the analyte.
　なお、演算部２Ｃが、第２のインジケータ１７Ｂが測定した水素イオン濃度（ｐＨ）等に応じて、第１のインジケータ１７Ａが測定したグルコース濃度を補正してもよい。なお、第１のインジケータ１７Ａと第２のインジケータ１７Ｂとが同じ蛍光体を異なる濃度で含んでいる場合も、測定したアナライトの濃度補正が可能である。 The computing unit 2C may correct the glucose concentration measured by the first indicator 17A according to the hydrogen ion concentration (pH) measured by the second indicator 17B. Even when the first indicator 17A and the second indicator 17B contain the same phosphor at different concentrations, the concentration of the measured analyte can be corrected.
　測定するアナライトに応じて、インジケータ１７の種類は選択される。さらに、３個以上の凹部のある蛍光センサが、それぞれの凹部に異なるインジケータを有していてもよい。インジケータとしては、例えば、カリウムイオンを測定する場合には蛍光残基を有するクラウンエーテル誘導体などを含む蛍光体（蛍光波長：５００ｎｍ）を用いることができ、酸素を測定する場合には、トリス（４，７－ジフェニル－１，１０－フェナントロリン）過塩素酸ルテニウム（ＩＩ）分子を含む蛍光体（蛍光波長：５３０ｎｍ）を用いることができる。 The type of indicator 17 is selected according to the analyte to be measured. Further, a fluorescent sensor having three or more recesses may have a different indicator in each recess. As the indicator, for example, when measuring potassium ion, a phosphor (fluorescence wavelength: 500 nm) including a crown ether derivative having a fluorescent residue can be used, and when measuring oxygen, Tris (4 , 7-diphenyl-1,10-phenanthroline) a phosphor containing a ruthenium (II) perchlorate molecule (fluorescence wavelength: 530 nm) can be used.
　マルチセンシング型の蛍光センサ４Ｅは蛍光センサ４等の効果を有し、さらに、１つのセンサで複数のアナライトを測定できるため、より環境や生体情報を正確に計測可能である。さらに測定したアナライト濃度の補正が可能であるため、より高精度である。 The multi-sensing type fluorescence sensor 4E has the effect of the fluorescence sensor 4 and the like, and moreover, since a plurality of analytes can be measured with one sensor, the environment and biological information can be measured more accurately. Furthermore, since the measured analyte concentration can be corrected, the accuracy is higher.
　測定するアナライトの種類、すなわちインジケータの種類により、照射する励起光の波長を変えてもよい。すなわち、第１のインジケータ１７Ａに第１の波長の励起光Ｅ１を照射し、第２のインジケータ１７Ｂには第２の波長の励起光Ｅ２が照射されてもよい。例えば、フェニルボロン酸誘導体を蛍光体として含む第１のインジケータ１７Ａには波長３７５ｎｍの第１の励起光Ｅ１を照射し、ＧＦＰ（green fluorescent protein）誘導体を蛍光体として含む第２のインジケータ１７Ｂには波長４９０ｎｍの第２の励起光Ｅ２を照射してもよい。 The wavelength of the excitation light to be irradiated may be changed depending on the type of analyte to be measured, that is, the type of indicator. That is, the first indicator 17A may be irradiated with the excitation light E1 having the first wavelength, and the second indicator 17B may be irradiated with the excitation light E2 having the second wavelength. For example, the first indicator 17A including a phenylboronic acid derivative as a phosphor is irradiated with a first excitation light E1 having a wavelength of 375 nm, and the second indicator 17B including a GFP (green fluorescent protein) derivative as a phosphor. You may irradiate the 2nd excitation light E2 of wavelength 490nm.
　異なる波長の励起光を照射するためには、蛍光センサ４Ｅのセンサ部１０Ｅは、第１の波長の励起光Ｅ１を発生する第１の発光素子であるＬＥＤ１５Ａ１と、第２の波長の励起光Ｅ２を発生する第２の発光素子であるＬＥＤ１５Ｂ１とを有していればよい。 In order to irradiate excitation light of different wavelengths, the sensor unit 10E of the fluorescence sensor 4E includes an LED 15A1 that is a first light emitting element that generates excitation light E1 of the first wavelength, and excitation light E2 of the second wavelength. LED15B1 which is the 2nd light emitting element which generate | occur | produces.
　または、図１２に示す変形例３の蛍光センサ４Ｆのセンサ部１０Ｆのように波長変換フィルタ８０を用いてもよい。例えば、ＬＥＤ１５Ｌが発生する波長３７５ｎｍの第１の波長の励起光Ｅ１を、第１のインジケータ１７Ａには、そのまま照射し、第２のインジケータ１７ＢにはＬＥＤ１５Ｌが発生する励起光Ｅ１を波長変換フィルタ８０により第２の波長である４６０ｎｍの励起光Ｅ２に変換して照射する。 Alternatively, the wavelength conversion filter 80 may be used as in the sensor unit 10F of the fluorescence sensor 4F of Modification 3 shown in FIG. For example, the wavelength conversion filter 80 emits the excitation light E1 of the first wavelength of 375 nm generated by the LED 15L to the first indicator 17A as it is, and the excitation light E1 generated by the LED 15L to the second indicator 17B. Is converted into excitation light E2 having a second wavelength of 460 nm and irradiated.
　また、図１３に示す変形例４の蛍光センサ４Ｇのセンサ部１０Ｇのようにインジケータ１７Ａ、１７Ｂが、第１のアナライト９Ａの量に応じた第１の蛍光Ｆ１を発生する第１の蛍光体８１Ａと第２のアナライト９Ｂの量に応じた第２の蛍光Ｆ２を発生する第２の蛍光体８１Ｂとを含む、同じものであってもよい。 Further, as in the sensor unit 10G of the fluorescence sensor 4G of the modification 4 shown in FIG. 13, the indicators 17A and 17B generate the first phosphor F1 that generates the first fluorescence F1 according to the amount of the first analyte 9A. 81A and the same thing including the 2nd fluorescent substance 81B which generate | occur | produces the 2nd fluorescence F2 according to the quantity of the 2nd analyte 9B may be sufficient.
　この場合には、第１の蛍光体８１Ａが発生する第１の蛍光Ｆ１を選択的に透過する第１のフィルタ１４Ａ１に覆われた第１のＰＤ素子１２Ａと、第２の蛍光体８１Ｂが発生する第２の蛍光Ｆ２を選択的に透過する第２のフィルタ１４Ｂ１に覆われた第２のＰＤ素子１２Ｂと、を有していれば、マルチセンシングが可能である。 In this case, the first PD element 12A covered by the first filter 14A1 that selectively transmits the first fluorescence F1 generated by the first phosphor 81A and the second phosphor 81B are generated. If it has 2nd PD element 12B covered with 2nd filter 14B1 which permeate | transmits the 2nd fluorescence F2 to perform selectively, multi sensing is possible.
　本発明は、上述した実施形態等に限定されるものではなく、本発明の要旨を変えない範囲において、種々の変更、改変等が可能である。 The present invention is not limited to the above-described embodiments and the like, and various changes and modifications can be made without departing from the scope of the present invention.
　本出願は、２０１２年４月２７日に日本国に出願された特願２０１２－１０３６０５号を優先権主張の基礎として出願するものであり、上記の開示内容は、本願明細書、請求の範囲、図面に引用されたものとする。 This application is filed on the basis of a priority claim based on Japanese Patent Application No. 2012-103605 filed in Japan on April 27, 2012. The above disclosure includes the present specification, claims, It shall be cited in the drawing.

Claims (8)

  1. 　主面に複数の凹部が形成されており、それぞれの凹部の壁面に蛍光を受光し検出信号を出力する光電変換素子が形成されている基板部と、
    　前記光電変換素子を覆う、励起光を遮断するフィルタと、
    　それぞれの前記凹部の内部に配設されている、前記励起光を受光するとアナライト量に応じた前記蛍光を発生するインジケータと、
    　前記凹部の開口を覆う、前記インジケータへの外光進入を遮断するが、アナライトは通過する遮光層と、
    　前記凹部の底面側から前記インジケータに励起光を照射する発光素子と、を具備することを特徴とする蛍光センサ。     A plurality of concave portions are formed on the main surface, and a substrate portion on which photoelectric conversion elements that receive fluorescence and output detection signals are formed on the wall surfaces of the respective concave portions,
    A filter that covers the photoelectric conversion element and blocks excitation light;
    An indicator that is arranged inside each of the recesses and generates the fluorescence according to the amount of analyte when receiving the excitation light;
    Covering the opening of the recess, blocking external light entry to the indicator, but the light-shielding layer through which the analyte passes,
    And a light emitting element that irradiates the indicator with excitation light from the bottom surface side of the recess.
  2. 　前記複数の凹部の底面に、それぞれ前記発光素子が配設されていることを特徴とする請求項１に記載の蛍光センサ。 The fluorescent sensor according to claim 1, wherein the light emitting elements are respectively disposed on the bottom surfaces of the plurality of recesses.
  3. 　前記基板部が、複数の発光素子が主面に配設された第１の基板部と、前記第１の基板部の前記主面と接合された複数の貫通孔が形成されている第２の基板部と、からなること特徴とする請求項２に記載の蛍光センサ。 The substrate part includes a first substrate part in which a plurality of light emitting elements are disposed on a main surface, and a plurality of through holes joined to the main surface of the first substrate part. The fluorescent sensor according to claim 2, further comprising a substrate portion.
  4. 　前記基板部の前記複数の凹部が貫通孔であり、前記複数の貫通孔の前記遮光層に覆われた開口と反対側の開口の直下領域を覆うように前記基板部に前記発光素子が接合されていることを特徴とする請求項１に記載の蛍光センサ。 The plurality of concave portions of the substrate portion are through holes, and the light emitting element is bonded to the substrate portion so as to cover a region directly below the opening of the plurality of through holes on the side opposite to the opening covered with the light shielding layer. The fluorescent sensor according to claim 1, wherein:
  5. 　前記複数のインジケータが、第１のアナライトの量に応じた第１の蛍光を発生する第１の蛍光体を含む第１のインジケータと、第２のアナライトの量に応じた第２の蛍光を発生する第２の蛍光体を含む第２のインジケータと、からなることを特徴とする請求項１から請求項４のいずれか１項に記載の蛍光センサ。 The plurality of indicators include a first indicator including a first phosphor that generates a first fluorescence according to the amount of the first analyte, and a second fluorescence according to the amount of the second analyte. The fluorescent sensor according to any one of claims 1 to 4, further comprising a second indicator including a second phosphor that generates light.
  6. 　前記第１のインジケータに第１の波長の励起光が照射され、前記第２のインジケータに第２の波長の励起光が照射されることを特徴とする請求項５に記載の蛍光センサ。 6. The fluorescence sensor according to claim 5, wherein the first indicator is irradiated with excitation light having a first wavelength, and the second indicator is irradiated with excitation light having a second wavelength.
  7. 　前記インジケータが、第１のアナライトの量に応じた第１の蛍光を発生する第１の蛍光体と第２のアナライトの量に応じた第２の蛍光を発生する第２の蛍光体とを含み、
    　前記第１の蛍光体が発生する第１の蛍光を透過する第１のフィルタに覆われた第１の光電変換素子と、前記第２の蛍光体が発生する第２の蛍光を透過する第２のフィルタに覆われた第２の光電変換素子と、を有することを特徴とする請求項１から請求項４のいずれか１項に記載の蛍光センサ。     A first phosphor that generates a first fluorescence according to an amount of the first analyte, and a second phosphor that generates a second fluorescence according to an amount of the second analyte; Including
    The first photoelectric conversion element covered with the first filter that transmits the first fluorescence generated by the first phosphor, and the second that transmits the second fluorescence generated by the second phosphor. The fluorescent sensor according to claim 1, further comprising: a second photoelectric conversion element covered with a filter.
  8. 　前記基板部、前記フィルタ、前記インジケータ、前記遮光層および前記発光素子を含むセンサ部を有する針先端部と、体外に配置される本体部の嵌合部と嵌合するコネクタ部と、を具備する、体内のアナライトを計測する針型センサであることを特徴とする請求項１に記載の蛍光センサ。 A needle tip portion having a sensor portion including the substrate portion, the filter, the indicator, the light shielding layer, and the light emitting element; and a connector portion fitted to a fitting portion of a main body portion arranged outside the body. The fluorescent sensor according to claim 1, wherein the fluorescent sensor is a needle-type sensor that measures an analyte in the body.
PCT/JP2013/062368 2012-04-27 2013-04-26 Fluorescence sensor WO2013161991A1 (en)

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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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