CN110243793A - A kind of chemiluminescence optical fiber biosensor - Google Patents
A kind of chemiluminescence optical fiber biosensor Download PDFInfo
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- CN110243793A CN110243793A CN201910531191.7A CN201910531191A CN110243793A CN 110243793 A CN110243793 A CN 110243793A CN 201910531191 A CN201910531191 A CN 201910531191A CN 110243793 A CN110243793 A CN 110243793A
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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
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Abstract
The present invention relates to a kind of chemiluminescence optical fiber biosensors characterized by comprising sample cell, for accommodating the determinand mixing sample and chemiluminescent substrate of enzyme label；Fibre-optical probe, the fibre-optical probe are fixed in the sample cell, and biological identification molecule is modified on the fibre-optical probe, and the fibre-optical probe is used to acquire the optical signal of determinand mixing sample chemiluminescence generation；Photodetector, the photodetector connect the fibre-optical probe, and the photodetector is used to the fluorescence signal of acquisition being converted to electric signal；Control system, the control system are electrically connected the multiposition valve, peristaltic pump and photodetector, and the present invention can be widely applied in environmental monitoring technology field.
Description
Technical field
The present invention relates to a kind of optical fiber biosensors to belong to especially with regard to a kind of chemiluminescence optical fiber biosensor
In environmental monitoring technology field.
Background technique
In today that environmental pollution is got worse, environmental monitoring is indispensable step for environmental pollution improvement
Suddenly, it can scientifically measure each factor for influencing environmental quality.The frequent generation of water environment pollution event, to environment
The requirement it is also proposed that new is detected, the on-site test in situ based on portable instrumentation becomes the new developing direction of environment measuring.
The monitoring of toxic pollutant still relies primarily on traditional Laboratory Instruments analysis method, such as gas phase color in water environment at present
Spectrum, high performance liquid chromatography etc., these instruments have high sensitivity, testing result accurate etc. excellent the detection of environmental contaminants
Point.
However, traditional instrument detection method must rely on complicated and expensive instrument, and quantitative detection is carried out to substance
Operating process it is cumbersome, testing cost is high, it is difficult to meet real-time detection in situ required by environment measuring, therefore, be based on water
Environmental Pollution is puted forth effort to research and develop and production high sensitivity, high specificity, pollutant monitoring instrument easy to operate and portable exists
Environmental monitoring technology is of great practical significance in field.
Summary of the invention
In view of the above-mentioned problems, the object of the present invention is to provide a kind of high sensitivities, high specificity, easy to operate and portable
Chemiluminescence optical fiber biosensor.
To achieve the above object, the present invention takes following technical scheme: a kind of chemiluminescence optical fiber biosensor, special
Sign is, comprising: sample cell, for accommodating the determinand mixing sample and chemiluminescent substrate of enzyme label；Fibre-optical probe, it is described
Fibre-optical probe is fixed in the sample cell, and biological identification molecule, the fibre-optical probe are modified on the fibre-optical probe
For acquiring the optical signal of determinand mixing sample chemiluminescence generation；Photodetector, described in photodetector connection
Fibre-optical probe, the photodetector are used to the fluorescence signal of acquisition being converted to electric signal；Control system, the control system
It is electrically connected the multiposition valve, peristaltic pump and photodetector.
Preferably, the inlet of the sample cell is provided with multiposition valve, and the entrance passes through the multiposition valve to the sample
The to be measured of chemiluminescent substrate, buffer PBS, antibody and standard items mixed liquor, regenerated liquid SDS and enzyme label is passed through in product pond
Object.
Preferably, the exit of the sample cell is provided with peristaltic pump.
Preferably, the fibre-optical probe is removal by the coat of optical fiber surface, and it is molten that optical fiber is immersed to the Piranha prepared
It is impregnated in liquid, remove the organic matter on optical fiber and completes the fibre-optical probe that optical fiber surface hydroxylating obtains.
Preferably, the fibre-optical probe is popped one's head in using silica fibre.
Preferably, the refractive index of the silica fibre probe fibre core is 1.456, and numerical aperture 0.22, core diameter is 600 μ
m；When being put into solution after the silica clad removal that the silica fibre is popped one's head in, the refractive index of solution is 1.33 meters, and solution passes
The modulus for broadcasting light is 1758.
Preferably, the control system includes data processor and computer, and the data processor is used for the light
The electric signal of electric explorer conversion is sent to the computer, is provided with parameter setting module in the computer, for preparatory
Set running parameter；Wriggling pump control module, for controlling unlatching or the pass of the peristaltic pump according to preset running parameter
It closes；Multiposition valve control module, for controlling opening or closing for the multiposition valve according to preset running parameter；Data analysis
Module, for the electric signal according to predetermined quantitative measurement standard curve and data processor transmission, analysis is obtained
Mix the concentration of determinand in sample；Data memory module, for storing the electric signal and divide that the data processor is sent
Analyse obtained testing concentration.
Preferably, the fibre-optical probe is fixedly connected using optical fiber connector with the sample cell.
Preferably, the photodetector uses photodiode.
The invention adopts the above technical scheme, which has the following advantages: the present invention is based on chemiluminescence and optical fiber are complete
Principles of internal reflection, chemiluminescent process progresses are not necessarily to excitation light source, and optical fiber biosensor structure is relatively simple, and the two combines design
Biosensor be easily achieved miniaturization, material absorbing in reaction system chemically reacts discharged energy, excites electronics
Excitation state is transitted to, and releases energy return to ground state quickly, generates chemiluminescence phenomenon.Optical fiber as light signal transduction element,
When the optical signal that chemiluminescence generates enters optical fiber at a proper angle, can be propagated in light with total reflection mode, and pass
It is handed at photodetector, detects the size of optical signal.Chemiluminescence usually requires to carry out under the catalytic action of catalyst,
It is connected to the biosensor on fibre-optical probe by absorption or biological compatible reaction, when biosensor contacts chemistry
When luminescence-producing reaction substrate, the generation of meeting cataluminescence reaction generates chemiluminescence signal.By the change for measuring optical signal
Change, the change information of biosensor can be found out.Namely the case where biosensor and analyte interaction, chemistry
Luminous signal intensity and the concentration of biological substance have quantitative relationship, so as to realize the quantitative detection to biological substance, tool
There is high sensitivity, high specificity, easy to operate and portable, can be widely applied in environmental monitoring technology field.
Detailed description of the invention
Fig. 1 is overall structure diagram of the invention；
Fig. 2 is the structural schematic diagram of fibre-optical probe in the present invention.
Specific embodiment
Come to carry out detailed description to the present invention below in conjunction with attached drawing.It should be appreciated, however, that attached 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 1, chemiluminescence optical fiber biosensor provided by the invention includes 1, six valve 2 of sample cell, wriggles
Pump 3, fibre-optical probe 4, photodetector 5 and control system 6, wherein be modified with biological identification molecule on fibre-optical probe 4.
The inlet of sample cell 1 is provided with six valves 2, and entrance is used to be passed through chemical hair into sample cell 1 by six valves 2
Light substrate, buffer PBS (phosphate buffer), antibody and standard items mixed liquor, regenerated liquid SDS (anion scale remover) and enzyme
The determinand (such as HRP enzyme, that is, horseradish peroxidase-labeled goat anti-mouse antibody determinand) of label.The outlet of sample cell 1
Place is provided with peristaltic pump 3, exports and sample cell 1 is discharged for the waste liquid after will test.Fibre-optical probe 4, light are provided in sample cell 1
4 connection photodetector 5 of fibre probe, fibre-optical probe 4 are used to acquire the optical signal of determinand mixing sample chemiluminescence generation, light
Electric explorer 5 is used to the fluorescence signal of acquisition being converted to electric signal.Six valves 2, peristaltic pump 3 and photodetector 5 are also distinguished
Be electrically connected connected control system 6, and control system 6 is used to be analyzed and processed to obtain to electric signal the concentration of determinand in mixing sample.
In a preferred embodiment, fibre-optical probe 4 is that directly optical fiber is immersed the coat of optical fiber surface by removal
V (dense H2SO4): v (H2O2It is impregnated in)=3:1 mixed liquor, that is, piraha (Piranha solution) solution, removes the organic matter on optical fiber
And complete what optical fiber surface hydroxylating obtained, so that the modification and being capable of fixing of biomolecule of fibre-optical probe 4.
In a preferred embodiment, as shown in Fig. 2, fibre-optical probe 4 can be using silica fibre probe 4, quartzy light
The refractive index of fibre 4 fibre cores of probe is 1.456, and numerical aperture 0.22, core diameter is 600 μm.When by silica fibre probe 4 quartz
When being put into solution after covering removal, the refractive index of solution is 1.33 meters, and the modulus that solution propagates light is 1758.
In a preferred embodiment, fibre-optical probe 4 can be modified with different function group (example according to different demands
Such as hydroxyl or amino) biological identification molecule can replace after having detected certain determinand mixing sample and be modified with other lifes
Object identifies the fibre-optical probe 4 of molecule, to realize the detection of various samples.
In a preferred embodiment, fixation of the fibre-optical probe 4 in sample cell 1 can be using optical fiber connector for example
FC has the advantages that easy to disassemble.
In a preferred embodiment, photodetector 5 can use photodiode.
In a preferred embodiment, control system 6 includes data processor 61 and computer 62, data processor 61
Electric signal for converting photodetector 5 is sent to computer 62, and parameter setting module is provided in computer 62, is wriggled
Pump control module, six valve control modules, data analysis module, data memory modules.
Parameter setting module is for presetting running parameter, wherein running parameter includes instrument serial ports and baud number etc.
(such as serial port setting is 5,9600) baud number is set as.Wriggling pump control module is used for according to preset running parameter, control
Peristaltic pump 3 opens or closes.Six valve control modules be used to be controlled according to preset running parameter six valves 2 unlatching or
It closes.Data analysis module is used for the telecommunications sent according to predetermined quantitative measurement standard curve and data processor 61
Number, analysis obtains the concentration of determinand in mixing sample, realizes the quantitative detection of determinand.Data memory module is for storing number
The testing concentration etc. that the electric signal and analysis sent according to processor 61 obtains.
Below by the application method of the specific embodiment chemiluminescence optical fiber biosensor that the present invention will be described in detail:
1) for fibre-optical probe 4 using silica fibre probe 4, refractive index 1.456, photodetector 5 uses photodiode,
Bifunctional group is connected with silanization process by hydroxylating on silica fibre probe 4, the bifunctional group other end connects MC-
LR-OVA conjugate (microcapsule algae toxin-ovalbumin conjugate) constitutes biological identification molecule.
2) silica fibre modified probe 4 is fixed in sample cell 1 by optical fiber connector.
3) computer 62 presets running parameter, according to preset running parameter, controls peristaltic pump 3 and six valves 2 open
It is dynamic, anti-MC-LR antibody (microcapsule algae toxin) is passed through into sample cell 1, after reacting a period of time, PBS washes away extra free anti-
Body.
4) it is passed through the secondary antibody of the label HRP enzyme mixed with MC-LR standard solution into sample cell 1, makes it and is connected to stone
Antibody on English fibre-optical probe 4 is specifically bound.
5) chemiluminescent substrate is passed through into sample cell 1, chemiluminescent substrate is anti-by HRP enzymatic generation chemiluminescence
It answers, generates chemiluminescence signal.
6) chemiluminescence signal that 4 acquisition of silica fibre probe generates is passed to photodetector 5, and optical signal is converted into telecommunications
Number.
7) electric signal that photodetector 5 converts is sent to computer 62 by data processor 61.
8) electric signal that computer 62 is sent according to the quantitative measurement standard curve and data processor 61 that determine early period, point
Analysis obtains the concentration of determinand in mixing sample, realizes the quantitative detection to determinand.
The various embodiments described above are merely to illustrate the present invention, wherein the structure of each component, connection type and manufacture craft etc. are all
It can be varied, all equivalents and improvement carried out based on the technical solution of the present invention should not exclude
Except protection scope of the present invention.
Claims (9)
1. a kind of chemiluminescence optical fiber biosensor characterized by comprising
Sample cell, for accommodating the determinand mixing sample and chemiluminescent substrate of enzyme label；
Fibre-optical probe, the fibre-optical probe are fixed in the sample cell, are modified with bio-identification on the fibre-optical probe
Molecule, the fibre-optical probe are used to acquire the optical signal of determinand mixing sample chemiluminescence generation；
Photodetector, the photodetector connect the fibre-optical probe, and the photodetector is used for the fluorescence that will be acquired
Signal is converted to electric signal；
Control system, the control system are electrically connected the multiposition valve, peristaltic pump and photodetector.
2. a kind of chemiluminescence optical fiber biosensor as described in claim 1, which is characterized in that the entrance of the sample cell
Place is provided with multiposition valve, and the entrance is passed through chemiluminescent substrate, buffer into the sample cell by the multiposition valve
The determinand of PBS, antibody and standard items mixed liquor, regenerated liquid SDS and enzyme label.
3. a kind of chemiluminescence optical fiber biosensor as described in claim 1, which is characterized in that the outlet of the sample cell
Place is provided with peristaltic pump.
4. a kind of chemiluminescence optical fiber biosensor as described in claim 1, which is characterized in that the fibre-optical probe is to go
Except optical fiber being immersed in the Piranha solution prepared and is impregnated by the coat of optical fiber surface, the organic matter on removal optical fiber is simultaneously complete
The fibre-optical probe obtained at optical fiber surface hydroxylating.
5. a kind of chemiluminescence optical fiber biosensor as described in claim 1, which is characterized in that the fibre-optical probe uses
Silica fibre probe.
6. a kind of chemiluminescence optical fiber biosensor as claimed in claim 5, which is characterized in that the silica fibre probe
The refractive index of fibre core is 1.456, numerical aperture 0.22, and core diameter is 600 μm；When the silica clad that the silica fibre is popped one's head in
When being put into solution after removal, the refractive index of solution is 1.33 meters, and the modulus that solution propagates light is 1758.
7. a kind of chemiluminescence optical fiber biosensor as claimed in claim 2, which is characterized in that the control system includes
Data processor and computer, electric signal of the data processor by converting the photodetector be sent to it is described based on
Calculation machine is provided in the computer
Parameter setting module, for presetting running parameter；
Wriggling pump control module, for controlling opening or closing for the peristaltic pump according to preset running parameter；
Multiposition valve control module, for controlling opening or closing for the multiposition valve according to preset running parameter；
Data analysis module, for the telecommunications according to predetermined quantitative measurement standard curve and data processor transmission
Number, analysis obtains the concentration of determinand in mixing sample；
Data memory module, for storing the electric signal that the data processor is sent and the testing concentration that analysis obtains.
8. a kind of chemiluminescence optical fiber biosensor as described in any one of claim 1 to 7, which is characterized in that the light
Fibre probe is fixedly connected using optical fiber connector with the sample cell.
9. a kind of chemiluminescence optical fiber biosensor as described in any one of claim 1 to 7, which is characterized in that the light
Electric explorer uses photodiode.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
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| CN201910531191.7A CN110243793A (en) | 2019-06-19 | 2019-06-19 | A kind of chemiluminescence optical fiber biosensor |
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| Application Number | Priority Date | Filing Date | Title |
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| CN201910531191.7A CN110243793A (en) | 2019-06-19 | 2019-06-19 | A kind of chemiluminescence optical fiber biosensor |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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 |
| CN112504986A (en) * | 2021-02-05 | 2021-03-16 | 成都益清源科技有限公司 | Optical probe for effectively improving full-spectrum signal quality |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101246119A (en) * | 2007-03-23 | 2008-08-20 | 国家纳米技术与工程研究院 | Optical fiber biological sensing system |
| CN109312287A (en) * | 2016-07-29 | 2019-02-05 | 休斯顿大学系统 | The system and method for detecting chemiluminescence reaction |
-
2019
- 2019-06-19 CN CN201910531191.7A patent/CN110243793A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101246119A (en) * | 2007-03-23 | 2008-08-20 | 国家纳米技术与工程研究院 | Optical fiber biological sensing system |
| CN109312287A (en) * | 2016-07-29 | 2019-02-05 | 休斯顿大学系统 | The system and method for detecting chemiluminescence reaction |
Non-Patent Citations (1)
| Title |
|---|
| RONG YANG，ET AL.: "Development of novel portable and reusable fiber optical chemiluminescent biosensor and its application for sensitive detection of microcystin-LR", 《BIOSENSORS AND BIOELECTRONICS》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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 |
| CN112504986A (en) * | 2021-02-05 | 2021-03-16 | 成都益清源科技有限公司 | Optical probe for effectively improving full-spectrum signal quality |
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Application publication date: 20190917 |
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