CN105842218A - Nanometer microfluidic accurate and quantitative fluoroimmunoassay detection platform - Google Patents
Nanometer microfluidic accurate and quantitative fluoroimmunoassay detection platform Download PDFInfo
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- CN105842218A CN105842218A CN201610346755.6A CN201610346755A CN105842218A CN 105842218 A CN105842218 A CN 105842218A CN 201610346755 A CN201610346755 A CN 201610346755A CN 105842218 A CN105842218 A CN 105842218A
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6402—Atomic fluorescence; Laser induced fluorescence
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/531—Production of immunochemical test materials
- G01N33/532—Production of labelled immunochemicals
- G01N33/533—Production of labelled immunochemicals with fluorescent label
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2201/00—Features of devices classified in G01N21/00
- G01N2201/06—Illumination; Optics
- G01N2201/061—Sources
- G01N2201/06113—Coherent sources; lasers
- G01N2201/0612—Laser diodes
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Abstract
The invention discloses a nanometer microfluidic accurate and quantitative fluoroimmunoassay detection platform, which consists of two parts, namely a machine body and a machine main body, wherein the machine body refers to an external structure of the platform, and comprises a housing, a power jack, a touch display screen, a reagent card insertion opening, a printed information output port and an external communication interface; the machine main body refers to an internal structure of the platform, and comprises a power unit, a microcomputer, a micro-printer, an MPPC photon counter, a laser source, an optical unit, an electronic quality control unit and a consumable position control and reagent identification module. According to the platform, the laser source is adopted, and can be controlled by the intelligent control technology to output exciting laser which adapts to the characteristics of detected samples and of which the wavelength and power are constant according to different detected samples; with adoption of the MPPC photon counter, weak light signal detection sensitivity and photon counting accuracy are greatly improved.
Description
Technical field
The invention belongs to photodetection and automatic control technology field, be specifically related to a kind of nanometer micro-fluidic accurate quantification fluorescence immunoassay detection platform.
Background technology
MPPC is the product that one is referred to as silicon photoelectric multiplier (SiPM, silicon PM), the photon counting product being made up of multiple APD pixels being operated in Geiger pattern.Although MPPC is a kind of optoelectronic semiconductor essentially, but it has an outstanding photon counting ability, and is applicable to various needs and carries out among the application of pole low light level measurement.MPPC can be operated at lower voltages, is characterized in high-gain, high photon detection efficiency, high reaction speed, outstanding temporal resolution and wider spectral response range, and its performance has met the requirement of photon counting of higher levels.It addition, MPPC is also equipped with the specific advantage of the solid products such as insensitive, the anti-metal impact in magnetic field itself.
Cucumber is through the irradiation of ultraviolet light, after absorbing the incident light of certain wavelength, can launch the light that relatively lambda1-wavelength is slightly grown, when ultraviolet light once stops irradiating, and the light launched disappears the most therewith, and the light of this transmitting is referred to as fluorescence.Owing to the molecular structure of material is different, the wavelength that can absorb ultraviolet light and the wavelength launching fluorescence are the most different, utilize this characteristic can carry out qualitative to test substance.Under certain condition, test substance concentration is the highest, and the fluorescence that ultraviolet light is launched after irradiating is the strongest；Otherwise, concentration is the lowest, and the light launched is the most weak；Accordingly, test substance can be carried out quantitative analysis.
Fluoroimmunoassay technology is to utilize the high susceptibility of fluorescent technique to combine with the high degree of specificity of immunological technique, for immunology, clinical tissue chemistry and laboratory diagnosis provide an additive method can not substituted, there is the detection technique of individual style.The immunofluorence technic application history of existing nearly 60 years in medical science and biology.Along with development and the application of immunofluorescence technique, make it a kind of immunological technique conventional in microbiology, immunology, pathology and immunohistochemistry.
Fluorescence immunoassay detection technique has the advantages such as selectivity is strong, highly sensitive, practicality is good, therefore it is used for measuring the bioactive compound that content is the lowest, such as protein (enzyme, acceptor, antibody), hormone (steroid, thyroid hormone, phthalein hormone), medicine and microorganism etc..
Fluorescence immunity analyzer is a kind of important detection equipment in microbiology, immunology, pathology and immunohistochemistry.Fluorescence immunity analyzer is in addition to for the qualification of bacterium, virus, protozoon, worm and fungi etc. and the diagnosis of relevant disease at present, it is also widely used in the detection of serum antibody (including autoantibody), the diagnosis of autoimmune disease and research, the qualification of pathology antigen, antibody and complement and location, the pathological study of immune complex, antigenic relationship between bacterium, virus and host and acceptor, part research, the technology such as the research of the diagnosis of tumour immunity and research, Cell membrane antigens and acceptor thereof.
The accuracy of fluoroscopic examination result to be ensured, needs to ensure two testing conditions, and one is that the exciting light that light source sends is highly stable, including constant wavelength and constant power；Two is that photon detection instrument has the highest sensitivity and accuracy, i.e. also wants the faintest fluorescence to carry out photon counting accurately.
Existing fluorescence detection device is generally difficult to meet the two condition, so it is the most accurate to often lead to testing result.Its reason mainly includes two aspects: one is that existing fluorescence detection device generally uses LED light source, and photostability is poor, and exciting light is unstable, causes output resultant error big；Two is that existing fluorescence detection device generally uses photocell or array image sensor as fluorescence detection equipment, and detectivity and photon counting accuracy for faint optical signal are the most undesirable, cause testing result not accurate enough.
Summary of the invention
Unstable and for the low problem of weak optical signal detection sensitivity in order to solve excitation source that existing fluorescence detection device exists, the present invention provides a kind of nanometer micro-fluidic accurate quantification fluorescence immunoassay detection platform, described platform uses LASER Light Source, and pass through intelligent control technology so that the exciting light with constant wavelength and power that light source can adapt with its feature according to the different outputs of tested sample；Use MPPC as photon counter, detectivity for faint optical signal and photon counting accuracy are greatly improved.
For realizing above-mentioned target, the present invention by the following technical solutions:
A kind of nanometer micro-fluidic accurate quantification fluorescence immunoassay detection platform, is made up of fuselage and body two parts.Described fuselage refers to the external structure of platform, including outer cover, outlet, touch display screen, reagent card insert port, type information delivery outlet and external communication interface.Described body refers to the internal structure of platform, including power subsystem, microcomputer, mini-printer, MPPC photon counter, LASER Light Source, optical unit, electronics Quality Control unit, consumptive material position control and reagent information identification module.The input of described power subsystem is connected with the outlet of described fuselage, and output is connected with all parts of described body, provides power supply for it.Described microcomputer is connected with other each parts in addition to optical unit in described body, receives the data of each parts, output order, controls each parts and performs corresponding operation.Described microcomputer is connected with the touch display screen of described fuselage, receives the instruction of user's input, display system work state information.Described consumptive material position control module is connected with the reagent card insert port of described fuselage.
Described power subsystem provides stable power supply for system, completely cuts off the external signal interference to native system.
Described microcomputer comprises processor, internal memory, I/O interface and operating system, there is provided interactive controlling and man-machine interface for system, complete the Row control of detection, result output, data storage and data interaction management function by the unit module within software coordinates simultaneously.
Described mini-printer is connected to internal microcomputer, is exported by testing result data-printing.
Described MPPC photon counter is the core cell of this platform, it is internally provided with temperature control unit, voltage control unit, gain control unit and simulation output interface, arranging high tension voltage, operating temperature point and output gain by digital interface, its counting statistics data are exported to rear end sampling unit by analog interface.
Described LASER Light Source is permanent wavelength perseverance luminous power output, and power output is adjustable, it is possible to obtain stable excitation source, for the fluorescence excitation of reagent.
Described LASER Light Source includes Wavelength control circuit and power control circuit, and described Wavelength control circuit receives the instruction that described microcomputer sends, and light source sends the wavelength constant of light in setting value；Described power control circuit receives the instruction that described microcomputer sends, and light source sends the power invariability of light in setting value.
Described optical unit includes reflecting, scatters, focuses on optical instrument, it is provided that the refraction of optical path signal, scatter, the function such as focusing.Laser emitting light projects reagent card detection zone by light path, and the fluorescence signal after exciting is returned in photon counter by light path again.
Described electronics Quality Control unit comprises photosignal automatic calibrator device, every time start or when operating between when reach setting cycle, electronics Quality Control unit can be activated, and automatically calibrates equipment, and automatically records calibration information.
This platform is scanned detection by consumptive material translation mode, and described consumptive material position control module comprises support holder structure, stepper motor and control system thereof, by stepper motor described in described Microcomputer control, drives described support holder structure traction reagent card to be scanned.
Described reagent information identification module uses Quick Response Code mode to encode reagent card information, and it is attached to the non-detection zone of reagent card, recognition unit in machine is by scanning Quick Response Code, extract the term of validity of reagent card, batch number, sequence number, Quality Control parameter and calibration data, for product control and the data analysis of detection.
The workflow of described detection platform comprises the following steps:
1) potent agent consumptive material is inserted described reagent card insert port；
2), after described consumptive material position control unit detects reagent card insertion, start stepper motor dragging reagent card and enter in platform lucifuge detecting position；
3) consumptive material load data is identified by described reagent information identification module, data is uploaded to microcomputer collection data later and calculates；
4) described microcomputer starts laser instrument, and the detection window district of reagent card is excited scanning, described MPPC photon counter programming count reagent card excite light quantity, and data are uploaded to described microcomputer；
5) described microcomputer is after getting the data of photon counter, the effective information that binding reagents card carries carries out result calculating, so that later stage inquiry in data being stored the database to the machine after completing to calculate, use described mini-printer result to be printed out simultaneously；
6) when platform is linked into external management system when, by described external communication interface, data are uploaded to Tomcat-AdminPortal.
Advantages of the present invention and having the beneficial effect that
1) with using photocell or array image sensor as compared with the prior art of core detection device, this platform photon counter based on MPPC, detectivity and accuracy for small-signal are substantially improved；
2) gain of MPPC photon counter can on-demand regulate, it is possible to the reagent card of dynamically adapting difference excitation intensity type；
3) present invention uses laser diode to substitute common light emitting diode as light source, and passes through dynamic regulation so that emergent light wavelength and power stability.
Accompanying drawing explanation
The invention will be further described with embodiment below in conjunction with the accompanying drawings.
Fig. 1 is the organism internal structure figure of a kind of nanometer of the present invention micro-fluidic accurate quantification fluorescence immunoassay detection platform.
Detailed description of the invention
Embodiment
See accompanying drawing 1, a kind of nanometer micro-fluidic accurate quantification fluorescence immunoassay detection platform, be made up of fuselage and body two parts.Fuselage refers to the external structure of platform, including outer cover, outlet, touch display screen, reagent card insert port, type information delivery outlet and external communication interface.Body refers to the internal structure of platform, including power subsystem, microcomputer, mini-printer, MPPC photon counter, LASER Light Source, optical unit, electronics Quality Control unit, consumptive material position control and reagent information identification module.The input of described power subsystem is connected with the outlet of described fuselage, and output is connected with all parts of described body, provides power supply for it.Described microcomputer is connected with other each parts in addition to optical unit in described body, receives the data of each parts, output order, controls each parts and performs corresponding operation.Described microcomputer is connected with the touch display screen of described fuselage, receives the instruction of user's input, display system work state information.Described consumptive material position control unit is connected with the reagent card insert port of described fuselage.
Power subsystem provides stable power supply for system, completely cuts off the external signal interference to native system.Power subsystem is the input of 12V direct current.
Microcomputer comprises processor, internal memory, I/O interface and operating system, there is provided interactive controlling and man-machine interface for system, complete the Row control of detection, result output, data storage and data interaction management function by the unit module within software coordinates simultaneously.Described processor uses Broadcom
BCM2837, operating system uses Linux.
Mini-printer uses thermal printer, is connected to internal microcomputer, is exported by testing result data-printing.
Temperature control unit, voltage control unit, gain control unit and simulation output interface it is provided with inside described MPPC photon counter, arranging high tension voltage, operating temperature point and output gain by digital interface, its counting statistics data are exported to rear end sampling unit by analog interface.
LASER Light Source uses laser diode, and described laser diode is permanent wavelength perseverance luminous power output, and power output is adjustable, it is possible to obtain stable excitation source, for the fluorescence excitation of reagent.
LASER Light Source includes Wavelength control circuit and power control circuit, and described Wavelength control circuit receives the instruction that described microcomputer sends, and light source sends the wavelength constant of light in setting value；Described power control circuit receives the instruction that described microcomputer sends, and light source sends the power invariability of light in setting value.
Optical unit includes reflecting, scatters, focuses on optical instrument, it is provided that the refraction of optical path signal, scatter, the function such as focusing.Laser emitting light projects reagent card detection zone by light path, and the fluorescence signal after exciting is returned in photon counter by light path again.
Electronics Quality Control unit comprises photosignal automatic calibrator device, every time start or when operating between when reach setting cycle, electronics Quality Control unit can be activated, and automatically calibrates equipment, and automatically records calibration information.
This platform is scanned detection by consumptive material translation mode, and described consumptive material position control module comprises support holder structure, stepper motor and control system thereof, by stepper motor described in described Microcomputer control, drives described support holder structure traction reagent card to be scanned.
Reagent information identification module uses Quick Response Code mode to encode reagent card information, and it is attached to the non-detection zone of reagent card, recognition unit in machine is by scanning Quick Response Code, extract the term of validity of reagent card, batch number, sequence number, Quality Control parameter and calibration data, for product control and the data analysis of detection.
External communication interface is provided with LAN, USB, WiFi interface module, can access LIS system (Laboratory Information by these interface modules
System, laboratory information system).
The workflow of detection platform comprises the following steps:
1) potent agent consumptive material is inserted reagent card insert port；
2), after consumptive material position control unit detects reagent card insertion, start stepper motor dragging reagent card and enter in platform lucifuge detecting position；
3) consumptive material load data is identified by reagent information identification module, data is uploaded to microcomputer collection data later and calculates；
4) microcomputer starts laser instrument, and the detection window district of reagent card is excited scanning, MPPC photon counter programming count reagent card excite light quantity, and data are uploaded to described microcomputer；
5) microcomputer is after getting the data of photon counter, the effective information that binding reagents card carries carries out result calculating, so that later stage inquiry in data being stored the database to the machine after completing to calculate, use described mini-printer result to be printed out simultaneously；
6) when platform is linked into external management system when, by described external communication interface, data are uploaded to Tomcat-AdminPortal.
It is last that it is noted that obviously above-described embodiment is only for clearly demonstrating example of the present invention, and not restriction to embodiment.For those of ordinary skill in the field, can also make other changes in different forms on the basis of the above description.Here without also cannot all of embodiment be given exhaustive.And the obvious change thus extended out or variation still in protection scope of the present invention among.
Claims (8)
1. a nanometer micro-fluidic accurate quantification fluorescence immunoassay detection platform, it is characterized in that: described detection platform is made up of fuselage and body two parts, described fuselage refers to the external structure of platform, including outer cover, outlet, touch display screen, reagent card insert port, type information delivery outlet and external communication interface；Described body refers to the internal structure of platform, including power subsystem, microcomputer, mini-printer, MPPC photon counter, LASER Light Source, optical unit, electronics Quality Control unit, consumptive material position control and reagent information identification module；The input of described power subsystem is connected with the outlet of described fuselage, and output is connected with all parts of described body, provides power supply for it；Described microcomputer is connected with other each parts in addition to optical unit in described body, receives the data of each parts, output order, controls each parts and performs corresponding operation；Described microcomputer is connected with the touch display screen of described fuselage, receives the instruction of user's input, display system work state information；Described consumptive material position control module is connected with the reagent card insert port of described fuselage.
2. a kind of nanometer micro-fluidic accurate quantification fluorescence immunoassay detection platform as claimed in claim 1, it is characterized in that: described MPPC photon counter is the core cell of described detection platform, it is internally provided with temperature control unit, voltage control unit, gain control unit and simulation output interface, arranging high tension voltage, operating temperature point and output gain by digital interface, its counting statistics data are exported to rear end sampling unit by analog interface.
3. a kind of nanometer micro-fluidic accurate quantification fluorescence immunoassay detection platform as claimed in claim 1, it is characterised in that: described LASER Light Source is permanent wavelength perseverance luminous power output, and power output is adjustable, it is possible to obtain stable excitation source, for the fluorescence excitation of reagent.
4. a kind of nanometer micro-fluidic accurate quantification fluorescence immunoassay detection platform as described in claim 1 or 3, it is characterized in that: described LASER Light Source includes Wavelength control circuit and power control circuit, described Wavelength control circuit receives the instruction that described microcomputer sends, and light source sends the wavelength constant of light in setting value；Described power control circuit receives the instruction that described microcomputer sends, and light source sends the power invariability of light in setting value.
5. a kind of nanometer micro-fluidic accurate quantification fluorescence immunoassay detection platform as claimed in claim 1, it is characterized in that: described electronics Quality Control unit comprises photosignal automatic calibrator device, every time start or when operating between when reach setting cycle, electronics Quality Control unit is activated, equipment is calibrated automatically, and automatically records calibration information.
6. a kind of nanometer micro-fluidic accurate quantification fluorescence immunoassay detection platform as claimed in claim 1, it is characterized in that: described detection platform is scanned detection by consumptive material translation mode, described consumptive material position control module comprises support holder structure, stepper motor and control system thereof, by stepper motor described in described Microcomputer control, described support holder structure traction reagent card is driven to be scanned.
7. a kind of nanometer micro-fluidic accurate quantification fluorescence immunoassay detection platform as claimed in claim 1, it is characterized in that: described reagent information identification module uses Quick Response Code mode to encode reagent card information, and it is attached to the non-detection zone of reagent card, recognition unit in machine is by scanning Quick Response Code, extract the term of validity of reagent card, batch number, sequence number, Quality Control parameter and calibration data, for product control and the data analysis of detection.
8. a kind of nanometer micro-fluidic accurate quantification fluorescence immunoassay detection platform as claimed in claim 1, it is characterised in that the workflow of described detection platform comprises the following steps:
1) potent agent consumptive material is inserted described reagent card insert port；
2), after described consumptive material position control unit detects reagent card insertion, start stepper motor dragging reagent card and enter in platform lucifuge detecting position；
3) consumptive material load data is identified by reagent information identification module, data is uploaded to microcomputer collection data later and calculates；
4) described microcomputer starts laser instrument, and the detection window district of reagent card is excited scanning, described MPPC photon counter programming count reagent card excite light quantity, and data are uploaded to described microcomputer；
5) described microcomputer is after getting the data of photon counter, the effective information that binding reagents card carries carries out result calculating, so that later stage inquiry in data being stored the database to the machine after completing to calculate, use described mini-printer result to be printed out simultaneously；
6) when platform is linked into external management system when, by described external communication interface, data are uploaded to Tomcat-AdminPortal.
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Cited By (2)
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| CN110672828A (en) * | 2019-09-10 | 2020-01-10 | 中国科学院上海技术物理研究所 | Rare animal blood detector of micro-fluidic laser immunity time domain spectrum |
| CN110672829A (en) * | 2019-09-10 | 2020-01-10 | 中国科学院上海技术物理研究所 | Microfluidic laser immunity time domain spectrum detection method |
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