CN106990086A - A kind of multichannel Evanescent Wave All-fiber biology sensor - Google Patents
A kind of multichannel Evanescent Wave All-fiber biology sensor Download PDFInfo
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- CN106990086A CN106990086A CN201710422048.5A CN201710422048A CN106990086A CN 106990086 A CN106990086 A CN 106990086A CN 201710422048 A CN201710422048 A CN 201710422048A CN 106990086 A CN106990086 A CN 106990086A
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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
- G01N21/6486—Measuring fluorescence of biological material, e.g. DNA, RNA, cells
Abstract
The present invention relates to a kind of multichannel Evanescent Wave All-fiber biology sensor, it is characterised in that the biology sensor includes Flow Sampling system, optical system and control circuit and signal processing system；It is divided into some Shu Guang after the laser alignment that semiconductor laser is sent, respective sample pond is transmitted into per light beam through corresponding all optical fiber system, each fibre-optical probe is through exciting generation fluorescence, the fluorescence signal of generation is transmitted into photodetector group after collecting filtering through all optical fiber system, the fluorescence signal on all fibre-optical probe surfaces of collection is converted into current signal by photodetector group, multichannel current signal is sent to computer after WeChat ID analog preamplifier and the processing of multichannel lock-in amplifier successively, realizes quantitative analysis while to sample multi objective；Computer also connects Collaborative Control module by parallel processing module, and parallel processing module also connects semiconductor laser, Collaborative Control wired in parallel connection peristaltic pump and ten valves by pulse signal generator.
Description
Technical field
The present invention is complete on a kind of multichannel evanescent wave based on total internal reflection evanescent wave principle and fluorescence analysis principle
Optical fiber biosensor.
Background technology
Environmental monitoring is as step indispensable in environmental protection work, and it can be to influenceing each factor of environmental quality
The measure of carry out science.At present, the trace toxic pollutant in water environment still lacks economical and practical analysis and detecting instrument and divided
Analysis method, currently mostly analyzes poisonous using traditional Laboratory Instruments analysis method (such as gas-chromatography, liquid chromatogram)
Specified risk material, and these instruments are typically only capable to be detected for a certain class material, so if needing to organic pollution, again
The toxicant of metal class and the class isomery of biotoxin class three detected, it is necessary to simultaneously equipment atomic absorption instrument, HPLC,
UPLC, ICP-MS (Inductively coupled plasma mass spectrometry) inductively coupled plasma matter
Spectrum, although these instruments have higher maturity and precision, but because the limitation of these detecting instrument sizes, it is impossible to
Detection, and expensive, complex operation in real time in situ is carried out, pretreatment is needed, this is not only for China's economic low developed area
For be difficult to, be also economically heavy burden to Chinese the eastern developed area.Therefore research and develop and production has certainly
The relatively inexpensive and easy to use of main intellectual property can detect the scientific instrument of multiple analytes in development fields of environmental monitoring in china simultaneously
There is important current demand in Instrument technology field.
Biology sensor according to total internal reflection fluorescent principle is biologic sensor for fast travelling waves of optical fibre mostly, in the prior art
Disclose single index optical fiber biosensor.Single index optical fiber biosensor structure is relatively simple, it is easy to accomplish miniaturization.Such as
Shown in Fig. 1, single index optical fiber biosensor principle is：When Ray Of Light enters optical fiber at a proper angle, understand to be totally reflected
Mode is propagated in a fiber, produces a kind of ripple for traversing optical fiber, and optical fiber is spread out of by the junction of optical fiber and other media, this
Ripple is a kind of energy wave with rapid decay, referred to as evanescent wave, the evanescent wave is propagated, and its transmission depth generally only has tens of to hundreds of
Nanometer.In the range of evanescent wave presence, the life on fibre-optical probe surface is connected to by absorption or with biological compatible reaction
Thing sensing element, when evanescent wave passes through biosensor, or generation optical signal (is such as marked with the biological substance of fluorescence molecule
It is excited and produces fluorescence), or cause evanescent wave with propagating intensity, phase or the frequency shift of light in optical fiber.Measure these
Change, you can obtain what the information of biosensor or optical fiber change, i.e. biosensor interacted with analyte
Information, when fluorescence molecule is excited, the concentration of this fluorescence intensity and biological substance has quantitative relation, so as to realize
Quantitative determination to biological substance, the Evanescent Wave All-fiber biology sensor according to this principle has sensitivity height, biospecific
Property is strong；The fast advantage of simple to operate, finding speed, but existing single index optical fiber biosensor can only once detect one
Index, is had some limitations using scope.
The content of the invention
In view of the above-mentioned problems, suddenly being died it is an object of the invention to provide a kind of multichannel that can realize multi objective Simultaneous Determination
Ripple all -fiber biology sensor.
To achieve the above object, the present invention takes following technical scheme：A kind of multichannel Evanescent Wave All-fiber bio-sensing
Device, it is characterised in that the biology sensor includes Flow Sampling system, optical system and control circuit and signal transacting system
System；The Flow Sampling system includes sample cell group, peristaltic pump and ten valves, and some sample cells in the sample cell group are successively
It is connected in series, one end of the sample cell group connects the peristaltic pump, and the other end of the sample cell group connects described ten valves,
A fibre-optical probe is fixedly installed in each sample cell, the different fibre-optical probes carry different rhetorical functions
Group, so that the different indexs in detectable same sample；The optical system includes semiconductor laser, based on single multimode light
The all optical fiber system and filter set of fine coupler；Control circuit and the signal processing system includes photodetector group, micro-
Signal imitation preamplifier, multichannel lock-in amplifier, parallel processing module, pulse signal generator, Collaborative Control module and
Computer；It is divided into some Shu Guang after the laser alignment that the semiconductor laser is sent, per light beam through corresponding all -fiber system
System is transmitted into the corresponding sample cell, and each fibre-optical probe is through exciting generation fluorescence, and the fluorescence signal of generation is through described complete
Fibre system is collected and the photodetector group is transmitted into after being filtered through the filter set, and the photodetector group will be received
The fluorescence signal on all fibre-optical probe surfaces of collection is converted into current signal, and multichannel current signal is successively through the WeChat ID
The computer is sent to after analog preamplifier and the processing of multichannel lock-in amplifier, it is fixed while realization to sample multi objective
Amount analysis；The computer also connects the Collaborative Control module, the parallel processing module by the parallel processing module
The semiconductor laser is also connected by the pulse signal generator, the Collaborative Control wired in parallel connection is described to wriggle
Pump and ten valves.
Preferably, the quantity for the sample cell that the sample cell group includes is four, and four sample cells are parallel longitudinal
It is spaced, the inner of four sample cells is provided with the connector for being fixedly connected with all optical fiber system, first
The inside top of sample cell and the 3rd sample cell is provided with sample export, and the inside bottom of the second sample cell and the 4th sample cell is set
It is equipped with sample inlet, the outer end of each sample cell is provided with connector, the import connection of the peristaltic pump described the
The sample export of one sample cell, the other end connection waste liquid pool of the peristaltic pump, the connector connection institute of first sample cell
The connector of the second sample cell is stated, the sample inlet of second sample cell connects the sample export of the 3rd sample cell, institute
The connector for stating the 3rd sample cell connects the connector of the 4th sample cell, and the sample inlet of the 4th sample cell passes through institute
State ten valves and be connected in parallel some samples.
Preferably, each all optical fiber system based on single multi-module optical fiber coupler includes a single-mode fiber, a list
Multi-module optical fiber coupler and two multimode fibres, one end of single multi-module optical fiber coupler are connected in parallel the single-mode fiber and
One multimode fibre, the other end of single multi-module optical fiber coupler connects the second multimode fibre, and exciting light is from the single-mode fiber
The fibre-optical probe, the fluorescence of the fibre-optical probe are entered by second multimode fibre through single multi-module optical fiber coupler
Signal enters the photoelectricity through single multi-module optical fiber coupler by second multimode fibre from first multimode fibre
Detector group.
Preferably, the WeChat ID analog preamplifier uses operational amplification circuit, will be defeated by the photodiode
Enter current signal direct proportion is converted into voltage signal.
The present invention is due to taking above technical scheme, and it has advantages below：1st, the present invention is put using four parallel intervals
The sample cell of fibre-optical probe is put and is equipped with, semiconductor laser is as excitation source, and exciting light is by optical splitter, based on single multimode
The all -fiber module of fiber coupler is transferred to fibre-optical probe, and different fibre-optical probes carries different modifying functional group, often
One fibre-optical probe produces fluorescence signal through exciting, by individually determine fluorescence signal can realize it is multinomial in a testing sample
The Simultaneous Determination of index；2nd, the present invention is provided with Flow Sampling system, and sample cell and testing sample are connected by ten valves, to be measured
Sampfe order is passed through multiple sample cells and Synchronization Analysis various ingredients, and sample introduction pipeline and program are simple, realize Full-automatic sequential
Flow Sampling.3rd, laser of the present invention by optical splitter during four fibre-optical probes carry out total internal reflection, in two-phase media
Interface produce evanescent wave, the biomolecule of evanescent wave excitation fiber detecting head surface fluorescence labeling produces fluorescence, single multimode light
Fine coupler is collected to fluorescence, converts optical signals to measurable electric signal by photodiode, and lock phase with multichannel
Amplifier will be detected that apparatus structure is simple, small volume, phosphor collection effect after faint measurable electric signal further amplification
Rate is high, ambient noise interference is small, can realize that multi objective is simultaneously and rapidly detected.To sum up, the present invention can be used for detection biology greatly
Molecule, organic molecule, virus and bacterium etc., it is adaptable to the technical field such as environmental monitoring, medical test, food Quality Control.
Brief description of the drawings
Fig. 1 is single index Evanescent Wave All-fiber biosensor structure schematic diagram in the prior art；
Fig. 2 is the multichannel Evanescent Wave All-fiber biosensor structure schematic diagram of the present invention；
Fig. 3 is all optical fiber system structural representation of the present invention；
Fig. 4 is the structural representation of the fibre-optical probe of the present invention.
Embodiment
Come to carry out the present invention detailed description below in conjunction with accompanying drawing.It should be appreciated, however, that accompanying drawing has been provided only more
Understand the present invention well, they should not be interpreted as limitation of the present invention.
As shown in Figure 2 and Figure 3, the multichannel Evanescent Wave All-fiber biology sensor that provides of the present invention, comprising optical system,
Flow Sampling system and control circuit and signal processing system.
Optical system include semiconductor laser 1, collimation lens 2, optical splitter 3, based on the complete of single multi-module optical fiber coupler
Fibre system 4, fibre-optical probe 5 and filter set 6；Wherein, all optical fiber system 4 includes single-mode fiber 41, single multimode fibre coupling
Device 42 and multimode fibre 43,44.
Flow Sampling system includes sample cell 7, peristaltic pump 8 and ten valves 9.
Circuit is controlled to include photodetector group 10, WeChat ID analog preamplifier 11, multichannel with signal processing system
Lock-in amplifier 12, computer 13, parallel processing module 14, Collaborative Control module 15 and pulse signal generator 16.
The Laser emission that semiconductor laser 1 is sent is to collimation lens 2, and laser shaping is directional light by collimation lens 2, accurate
Directional light after straight is sent to spectroscope 3 and is divided into some Shu Guang (embodiment of the present invention is four beams, as example, not limited to this),
Single-mode fiber 41 of the every light beam through the outgoing of spectroscope 3 through corresponding all optical fiber system 4 is incoming, and is coupled through single multimode fibre
Device 42 is passed to multimode fibre 44, and the fibre-optical probe 5 in sample cell 7 is transferred to through multimode fibre 44, is fixedly installed on each
The fluorescence molecule sample on the surface of fibre-optical probe 5 in sample cell 7 is through exciting generation fluorescence, and fluorescence signal is collected through multimode fibre 44
And filtered group 6 is transmitted into photodetector after filtering respectively after through single multi-module optical fiber coupler 42 being transmitted into multimode fibre 43
Group 10, filter set 10 is used to filter optical excitation signal, and photodetector group 10 is used for the surface of multiple fibre-optical probes 5 of collection
Different fluorescence signals be converted into current signal, multichannel current signal is locked through WeChat ID analog preamplifier 11 and multichannel successively
Computer 13 is sent to after the processing of phase amplifier 12, quantitative analysis while to sample multi objective is realized.
Computer 13 also connects Collaborative Control module 15 by parallel processing module 14, and parallel processing module 14 passes through pulse
Signal generator 16 connects semiconductor laser 1, and Collaborative Control module 15 is connected in parallel peristaltic pump 8 and ten valves 9, computer 13
Control parallel processing module 14 transmits a signal to Collaborative Control module 15, Collaborative Control module 15 according to the signal received so as to
Control the opening and closing of peristaltic pump 8 and ten valves 9.
Four sample cells 7 are provided with the embodiment of the present invention, four sample cells 7 are parallel longitudinal to be spaced, four samples
The inner in pond is provided with the connector for being fixedly connected with multimode fibre 43.The medial roof of first sample cell and the 3rd sample cell
Portion is provided with sample export, and the inside bottom of the second sample cell and the 4th sample cell is provided with sample inlet, each sample cell
Outer end is provided with connector, and the port of export connection waste liquid pool of peristaltic pump 8, the entrance point of peristaltic pump 8 connects the first sample cell
Sample export, the connector of the first sample cell connects the connector of the second sample cell, the sample inlet connection of the second sample cell
The sample export of 3rd sample cell, the connector of the 3rd sample cell connects the connector of the 4th sample cell, the sample of the 4th sample cell
Product entrance is connected in parallel some samples by ten valves 9.
In a preferred embodiment, four all optical fiber systems 4 based on single multi-module optical fiber coupler are connected by optical fiber
Connect the longitudinally fixed connection of device 17.
In a preferred embodiment, as shown in figure 3, each all optical fiber system 4 based on single multi-module optical fiber coupler
Include the single multimode fibre 43,44 of multi-module optical fiber coupler 42 and two of a single-mode fiber 41, one, single multi-module optical fiber coupler 42
One end is connected in parallel single-mode fiber 41 and multimode fibre 43, and the other end of single multi-module optical fiber coupler 42 connects another multimode fibre
44, exciting light enters fibre-optical probe 5 by single multi-module optical fiber coupler 43 from single-mode fiber 41 by multimode fibre 44, and optical fiber is visited
First 5 signals collected by multimode fibre 44 by single multi-module optical fiber coupler 42 from another root multimode fiber 43 be transmitted into
Enter photodetector group 10.
In a preferred embodiment, it is of the invention as shown in figure 4, fibre-optical probe 5 is the core component of whole system
Fibre-optical probe 5 uses silica fibre using tapered form, fibre-optical probe 5 is combined, and the refractive index of the fibre core of fibre-optical probe 5 is
1.456, numerical aperture is 0.22, and core diameter is 600 μm, molten when fibre-optical probe 5 is put into solution after silica clad is removed
The refractive index of liquid is 1.33, and its modulus for propagating light is 1758.The paint of black is coated in the end of non-sensitive part, laser is prevented
It is leaked in solution.Fibre-optical probe 5 passes through dismountable joints of optical fibre with the multimode fibre 44 in single multi-module optical fiber coupler
FC (FC) is attached.Use, realize a variety of while the present invention is by fibre-optical probe 5 of modifying different biological identification molecules
Detected while sample.
In a preferred embodiment, fibre-optical probe 5 can carry different modifying functional group (such as according to different demands
Hydroxyl or amino).The manufacturing process of fibre-optical probe 5 is：First by the optical fiber of 5.4cm length at the 2.4cm of even surface one end after
Coat is wiped off, is then placed in 30%HF solution immersion appropriate time, the diameter of optical fiber is corroded to 230~250 mesh, then will
The coat of optical fiber surface is all wiped off, makes optical fiber formation cone angle；Optical fiber is then immersed to the piraha solution newly prepared (dense
H2SO4:H2O2(v/v)=3:1) 60min is soaked, optical fiber is fully cleaned with ultra-pure water afterwards, aeration-drying is subsequently placed into
Case is dried, and is removed the organic matter on chip and is completed optical fiber surface hydroxylating, molten with 9800 μ L dry toluenes and 20 μ L APTES
Liquid；Then clean optical fiber is put into the solution and reacts 2h, the step silanization；By the optical fiber toluene solution after silanization
Cleaning three times, is then placed in and adds in 45 μ L DSC, 333.3 μ L DIEA, the solution of 9622 μ L toluene, reacts 2h, then uses toluene
Dried after rinse, this step connection bifunctional reagent；It will be connected to the optical fiber of bifunctional reagent alcohol flushing three times, then with height
Pure water rinsing is clean, is subsequently placed into aeration cabinet drying；Place the fiber in envelope antigen solution, at 4 DEG C overnight；Using
2mg/mL BSA closing chip 60min, to close non-specific adsorption sites, are then rinsed, N with high purity water2Drying, rear end
Face is dried with pitch-dark pitch-dark, standby.
In a preferred embodiment, filter set 6 by parallel multiple filter sets into each optical filter 6 is put
In the tail end of corresponding multimode fibre 43, to be filtered out to exciting light.
In a preferred embodiment, photodetector group 10 is made up of one group of photodiode, each pole of photoelectricity two
Fluorescence is converted into electric signal by pipe 10 close to the one side of optical filter.
In a preferred embodiment, WeChat ID analog preamplifier 11 uses operational amplification circuit, will be by photoelectricity
Detector input current signal direct proportion is converted into voltage signal.
Describe the use of the multichannel Evanescent Wave All-fiber biology sensor of the present invention in detail below by specific embodiment.
As shown in Fig. 2 the quantity of the sample cell of the present embodiment is 4, LASER Light Source uses wavelength for 635nm semiconductor
Laser, it is 15mW that pulsed signal generator 16, which is modulated into power output after the discontinuous light that frequency is 2800Hz, and collimation is saturating
The parallel circle hot spot that laser shaping is diameter about 1mm, fibre-optical probe 5 are made up mirror 2 of silica fibre, and refractive index is 1.456, glimmering
The multimode fibre that light is collected in all optical fiber system 4 based on single multi-module optical fiber coupler uses core diameter for 600 μm, and covering is
12.5, numerical aperture is 0.22, projection ratio ＜ 10 of the optical filter 6 to exciting light-7, to transmitance >=80% of fluorescence, photoelectricity is visited
Survey device and use photodiode.
During measurement, injection marked optical fiber in the biological sample solution of Cy5.5 fluorescent dyes, embodiment and visit in sample cell
The detectivity of first 5 couples of Cy5.5 fluorescence molecule solution is up to 10-9Mol/L, each interchannel signal amplitude and detectivity are poor
Different to be respectively less than 10%, the detection time for completing first order fluorescence signal is less than 20min, Atrazine and 2 can be detected simultaneously, 4-D is more
Individual index.
The various embodiments described above are merely to illustrate the present invention, wherein the structure of each part, connected mode and manufacture craft etc. are all
It can be varied from, every equivalents carried out on the basis of technical solution of the present invention and improvement should not be excluded
Outside protection scope of the present invention.
Claims (6)
1. a kind of multichannel Evanescent Wave All-fiber biology sensor, it is characterised in that the biology sensor includes Flow Sampling system
System, optical system and control circuit and signal processing system；The Flow Sampling system includes sample cell group, peristaltic pump and ten
Some sample cells in position valve, the sample cell group are sequentially connected in series, and one end of the sample cell group connects the peristaltic pump,
The other end of the sample cell group connects in described ten valves, each sample cell and is fixedly installed a fibre-optical probe, no
The same fibre-optical probe carries different rhetorical function groups；The optical system includes semiconductor laser, based on list
The all optical fiber system and filter set of multi-module optical fiber coupler；The control circuit includes photodetector with signal processing system
Group, WeChat ID analog preamplifier, multichannel lock-in amplifier, parallel processing module, pulse signal generator, Collaborative Control mould
Block and computer；
It is divided into some Shu Guang after the laser alignment that the semiconductor laser is sent, is sent out per light beam through corresponding all optical fiber system
The corresponding sample cell is mapped to, each fibre-optical probe is through exciting generation fluorescence, and the fluorescence signal of generation is through all -fiber
Systematic collection is transmitted into the photodetector group after simultaneously being filtered through the filter set, and the photodetector group is by collection
The fluorescence signal on all fibre-optical probe surfaces is converted into current signal, and multichannel current signal is simulated through the WeChat ID successively
The computer is sent to after preamplifier and the processing of multichannel lock-in amplifier, is quantitatively divided while realization to sample multi objective
Analysis；
The computer also connects the Collaborative Control module by the parallel processing module, and the parallel processing module also leads to
Cross the pulse signal generator and connect the semiconductor laser, the Collaborative Control wired in parallel connect the peristaltic pump and
Ten valves.
2. a kind of multichannel Evanescent Wave All-fiber biology sensor as claimed in claim 1, it is characterised in that the sample cell
The quantity for the sample cell that group includes is four, and four sample cells are parallel longitudinal to be spaced, four sample cells
The inner is provided with the medial roof of the connector for being fixedly connected with all optical fiber system, the first sample cell and the 3rd sample cell
Portion is provided with sample export, and the inside bottom of the second sample cell and the 4th sample cell is provided with sample inlet, each sample
The outer end in pond is provided with connector, and the import of the peristaltic pump connects the sample export of first sample cell, described compacted
The other end connection waste liquid pool of dynamic pump, the connector of first sample cell connects the connector of second sample cell, described
The sample inlet of second sample cell connects the sample export of the 3rd sample cell, the connector connection institute of the 3rd sample cell
The connector of the 4th sample cell is stated, the sample inlet of the 4th sample cell is connected in parallel some detected samples by described ten valves
Product.
3. a kind of multichannel Evanescent Wave All-fiber biology sensor as claimed in claim 1, it is characterised in that each base
Include a single-mode fiber, single a multi-module optical fiber coupler and two multimode light in all optical fiber system of single multi-module optical fiber coupler
Fibre, one end of single multi-module optical fiber coupler is connected in parallel the single-mode fiber and the first multimode fibre, single multimode light
The other end of fine coupler connects the second multimode fibre, and exciting light is logical through single multi-module optical fiber coupler from the single-mode fiber
Second multimode fibre is crossed into the fibre-optical probe, the fluorescence signal of the fibre-optical probe passes through second multimode fibre
Enter the photodetector group from first multimode fibre through single multi-module optical fiber coupler.
4. a kind of multichannel Evanescent Wave All-fiber biology sensor as described in any one of Claim 1-3, it is characterised in that
The filter set includes some optical filters, and each optical filter is arranged at the tail end of first multimode fibre, with right
Exciting light is filtered out.
5. a kind of multichannel Evanescent Wave All-fiber biology sensor as claimed in claim 4, it is characterised in that the photoelectricity is visited
Survey device group to be made up of some photodiodes, each photodiode is close to the corresponding optical filter.
6. a kind of multichannel Evanescent Wave All-fiber biology sensor as claimed in claim 5, it is characterised in that the WeChat ID
Analog preamplifier uses operational amplification circuit, will be converted into electricity by the photodiode input current signal direct proportion
Press signal.
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| CN110082334A (en) * | 2019-05-30 | 2019-08-02 | 华侨大学 | A kind of multichannel optical fiber fluorescent optical sensor |
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| CN110243795A (en) * | 2019-06-19 | 2019-09-17 | 中国人民大学 | A kind of evanescent wave Two Colour Fluorescence all -fiber biosensor |
| CN110425946A (en) * | 2019-08-07 | 2019-11-08 | 上海无线电设备研究所 | A kind of photodetector device |
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| CN113030033A (en) * | 2019-12-25 | 2021-06-25 | 中国科学院长春光学精密机械与物理研究所 | Fluorescence sensing system for biomarkers |
| CN111595830A (en) * | 2020-05-29 | 2020-08-28 | 迅检(义乌)科技有限公司 | Handheld dairy product risk pollutant detector and detection method |
| CN113804650A (en) * | 2021-09-18 | 2021-12-17 | 中国特种设备检测研究院 | Monitoring device and monitoring method |
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