CN204101452U - Based on the Streptavidin concentration sensor of single-mode fiber - Google Patents
Based on the Streptavidin concentration sensor of single-mode fiber Download PDFInfo
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- CN204101452U CN204101452U CN201420568366.4U CN201420568366U CN204101452U CN 204101452 U CN204101452 U CN 204101452U CN 201420568366 U CN201420568366 U CN 201420568366U CN 204101452 U CN204101452 U CN 204101452U
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- 108010090804 Streptavidin Proteins 0.000 title claims abstract description 32
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- 230000005540 biological transmission Effects 0.000 claims abstract description 23
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- 238000001179 sorption measurement Methods 0.000 claims description 9
- 238000001228 spectrum Methods 0.000 claims 2
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- 239000000243 solution Substances 0.000 description 16
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- 230000004048 modification Effects 0.000 description 3
- 238000012986 modification Methods 0.000 description 3
- 230000008569 process Effects 0.000 description 3
- WYTZZXDRDKSJID-UHFFFAOYSA-N (3-aminopropyl)triethoxysilane Chemical compound CCO[Si](OCC)(OCC)CCCN WYTZZXDRDKSJID-UHFFFAOYSA-N 0.000 description 2
- 108090001008 Avidin Proteins 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
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- 206010028980 Neoplasm Diseases 0.000 description 2
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- 238000010586 diagram Methods 0.000 description 2
- CWAFVXWRGIEBPL-UHFFFAOYSA-N ethoxysilane Chemical compound CCO[SiH3] CWAFVXWRGIEBPL-UHFFFAOYSA-N 0.000 description 2
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 2
- 238000007654 immersion Methods 0.000 description 2
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- 230000007246 mechanism Effects 0.000 description 2
- 108020004707 nucleic acids Proteins 0.000 description 2
- 102000039446 nucleic acids Human genes 0.000 description 2
- 150000007523 nucleic acids Chemical class 0.000 description 2
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- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 2
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- WGYKZJWCGVVSQN-UHFFFAOYSA-N propylamine Chemical group CCCN WGYKZJWCGVVSQN-UHFFFAOYSA-N 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 238000002310 reflectometry Methods 0.000 description 2
- YMXHPSHLTSZXKH-RVBZMBCESA-N (2,5-dioxopyrrolidin-1-yl) 5-[(3as,4s,6ar)-2-oxo-1,3,3a,4,6,6a-hexahydrothieno[3,4-d]imidazol-4-yl]pentanoate Chemical compound C([C@H]1[C@H]2NC(=O)N[C@H]2CS1)CCCC(=O)ON1C(=O)CCC1=O YMXHPSHLTSZXKH-RVBZMBCESA-N 0.000 description 1
- SNKZJIOFVMKAOJ-UHFFFAOYSA-N 3-Aminopropanesulfonate Chemical compound NCCCS(O)(=O)=O SNKZJIOFVMKAOJ-UHFFFAOYSA-N 0.000 description 1
- 102000035195 Peptidases Human genes 0.000 description 1
- 108091005804 Peptidases Proteins 0.000 description 1
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- 241001655322 Streptomycetales Species 0.000 description 1
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- 229960000789 guanidine hydrochloride Drugs 0.000 description 1
- PJJJBBJSCAKJQF-UHFFFAOYSA-N guanidinium chloride Chemical compound [Cl-].NC(N)=[NH2+] PJJJBBJSCAKJQF-UHFFFAOYSA-N 0.000 description 1
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- Investigating Or Analysing Materials By Optical Means (AREA)
Abstract
The utility model relates to a kind of Streptavidin concentration sensor based on single-mode fiber, it is characterized in that, described sensor is by light supply apparatus, spectroanalysis instrument, Transmission Fibers, three ports light rings, optical fiber stationary fixture, optical fiber probe and solution composition to be measured, the light signal output end of described light supply apparatus is connected by first port of Transmission Fibers with described three ports light rings, second port of described three port circulators is connected with described optical fiber probe by Transmission Fibers, described optical fiber probe is fixed by described optical fiber stationary fixture, place straight down and make its end immerse in solution to be measured completely, 3rd port of described three port circulators is connected with described spectroanalysis instrument by Transmission Fibers.Structure of the present utility model is simple, volume is little, safety non-toxic, can conveniently realize carrying out remote detection and real time on-line monitoring in vivo.
Description
Technical field
The utility model belongs to the crossing domain that optical fiber sensing technology is combined with bioprotein detection technique, be specifically related to a kind of Streptavidin concentration sensor based on single-mode fiber, utilize mechanism and the fiber end face Fresnel reflection principle of biotin and Streptavidin specific binding, realize the Concentration Testing of Streptavidin.
Background technology
Sensor refers to can experience certain measured signal, and converts thereof into the element of the physical quantitys such as electricity, sound, light, by sensing unit, and converting unit and transmission unit composition.The biologically active unit such as enzyme, antigen-antibody, nucleic acid are utilized to be referred to as biology sensor as the sensor of bio-sensing unit.Conventional biology sensor volume is large, needs a large amount of sample, and with poisonous (fluorescence) indicator, once can only do a kind of test, spended time is long, can not do in body and detect.
Optical fiber sensing technology take light wave as carrier, optical fiber is modulated in transmitting procedure for medium makes extraneous signal be converted to light signal, the physical features parameter of light signal such as light intensity (luminous power), wavelength, frequency, phase place and polarization state etc. are changed, carry out detection demodulation by the detector of receiving end again, the change of outer signals is embodied with the form of change in optical signal.Because optical fiber structure is stablized, be not subject to external environmental interference, be easily detected device and accept and detect, be therefore widely used in sensory field.
Optical fiber biosensor is using biologically active unit (enzyme, microorganism, animal vegetable tissue section, antigen-antibody, nucleic acid) as bio-sensing unit, utilize optical fiber as the reaction of signal converter captured target thing and responsive primitive, the extent of reaction is expressed by electric signal or light signal, thus obtains measured object concentration.This sensor can detect the change of biological respinse optical signal, by detecting the amount of the parameter determination analyte such as intensity, amplitude, phase place of light.Compared with other sensor, this sensor has by the interference of electromagnetic field, highly sensitive, quality is light, volume is little, be easy to multiplexing (networking), can the long-distance remote control advantage of monitoring.The application of optical fiber biosensor in life science, bioengineering, food industry, environment and agricultural etc., just at development, is especially applicable to remote measurement and on-line monitoring in biosome.
Streptavidin (Streptavidin, SA) is a kind of a kind of protein of purifying out from streptomycete, and molecular weight is 52.8k dalton.Because the chemical property that it is more favourable, so it can be used as the substitute of avidin.Streptavidin is a kind of tetramer, and be made up of four identical peptide chains, dissociation constant is 10
-14the mol/L order of magnitude, it and biotin (Biotin, also known as VB7 or biotin) have very strong suction-operated, and its every bar peptide chain can in conjunction with a biotin.The suction-operated of biotin and Streptavidin is one of current the strongest known noncovalent interaction, and both suction-operateds are species specific selective adsorptions, namely only have Avidin can only with Streptavidin generation specific adsorption, specific adsorption is there is not with other protein, there is selectivity, have very large potential in cancer cell, field of virus detection.In addition, Streptavidin-biotin system can effectively prevent organic solvent, denaturant (such as guanidine hydrochloride), washing agent and proteolytic enzyme from damaging it, also can retention properties stablize at extreme temperature and pH value, make Streptavidin be widely used in biomolecular science and bio-nanotechnology, the concentration measuring streptavidin is accurately vital.
Utilize general single mode fiber, end peels off overlay, cuts its end face flat and carries out surface silanization modification, recycle covalently bound mechanism, make surface carry biotin, form specific adsorption rete at fiber end face, thus mutually can adsorb to be formed to Streptavidin and stablize conjugate.The solution of streptavidin of variable concentrations is different from the situation that biotin combines, and the variations in refractive index of fiber end face is also different, and the light intensity of its end face Fresnel reflection is also different, can realize the measurement of solution of streptavidin concentration.
Summary of the invention
The purpose of this utility model is exactly: in order to solve conventional biosensor complex structure, bulky, sample requirements are large, utilize poisonous fluorescence indicator, the shortcoming that detection time is long, provides that a kind of structure is simple, volume is little, can realize the Streptavidin concentration sensor based on single-mode fiber of remote measurement and real time on-line monitoring in biosome.
The technical scheme that the utility model is taked for technical solution problem is:
A kind of Streptavidin concentration sensor based on single-mode fiber, it is characterized in that, described sensor is by light supply apparatus, spectroanalysis instrument, Transmission Fibers, three ports light rings, optical fiber stationary fixture, optical fiber probe and solution composition to be measured, the light signal output end of described light supply apparatus is connected by first port of Transmission Fibers with described three ports light rings, second port of described three port circulators is connected with described optical fiber probe by Transmission Fibers, described optical fiber probe is fixed by described optical fiber stationary fixture, place straight down and make its end immerse in solution to be measured completely, 3rd port of described three port circulators is connected with described spectroanalysis instrument by Transmission Fibers.
Described Transmission Fibers is single-mode fiber, and described optical fiber probe is that end is peelled off coat and cutting the single-mode fiber flat end face being modified with tool specific adsorption functional film layer.
The light that described light supply apparatus sends is from the first port input of three ports light rings, single-mode fiber is outputted to from the second port, the end of single-mode fiber is peelled off coat and is cut flat fiber end face, subsequently surface film modification is carried out to its end face, form optical fiber probe and absorption can be produced to Streptavidin, optical fiber probe for bio-sensing uses fixture to fix, place straight down and ensure that end is immersed in solution to be measured completely, end face Fresnel reflection light signal enters into spectroanalysis instrument via optical circulator from the 3rd port again, light signal is changed into concentration signal finally by process.
The beneficial effects of the utility model are:
1, for the general single mode fiber of transmitting optical signal, it is with low cost.
2, optical fiber probe for sensing is also be made by general single mode fiber, the process removing optical fiber coating and Qie Ping fiber end face can complete with ordinary optic fibre peeling plier and optical fiber cutter respectively, its manufacture craft is simple, be easy to realize, cheap, surface film modification technique also comparative maturity, it is large that optical fiber probe solves conventional biosensor volume, sample requirements is large, employing noxious material marks, the a series of shortcomings such as detection time is long, it is little that self also has volume, structure is simple, easy to operate, the advantages such as the on-line monitoring in biosome can be realized.
3, the principle by measuring Streptavidin concentration of analog bacterioprotein Concentration Testing can be applied to other material, if change the method for modifying of fiber end face, specificity adsorption selection can be produced to different materials, realize the detection of different material, such as to the selective enumeration method of cancer cell, testing process is made to become easier and convenient and practical.
Accompanying drawing explanation
Fig. 1 is the system architecture schematic diagram of the utility model embodiment.
Embodiment
Below in conjunction with accompanying drawing, embodiment of the present utility model is elaborated.
As shown in Figure 1, for the system architecture schematic diagram of the utility model embodiment, the Streptavidin concentration sensor based on single-mode fiber comprises light supply apparatus (10), spectroanalysis instrument (20), Transmission Fibers (30), three ports light rings (40), optical fiber stationary fixture (50), optical fiber probe (60) and solution to be measured (70).
The light signal that light supply apparatus (10) sends enters three ports light rings (40) by Transmission Fibers (30), and then arrive optical fiber probe (60) through another root Transmission Fibers (30), described optical fiber probe (60) detects solution to be measured (70), then optical signal feedback is carried out spectral analysis to spectroanalysis instrument (20).
General single mode fiber forms optical fiber probe (60) after processing as follows: first carry out pre-service to general single mode fiber: remove the coat of single-mode fiber tail end and cut planar end surface, with being immersed in the mixed solution of 30% hydrogen peroxide and 98% concentrated sulphuric acid after alcohol wipe, in solution, the volume ratio of two kinds of liquid is 1: 1, the immersion time is 30 minutes, makes fiber end face form silicone hydroxyl; Then 3-aminopropyl three ethoxy silane (APTES is immersed in, 3-APS), in the solution that absolute ethyl alcohol and deionized water mix, the volume ratio of three kinds of liquid is 2.5: 96.5: 1,3-aminopropyl three ethoxy silane is a kind of silane coupling agent, the hydroxyl covalent bond that the alkyl of its one end can produce with previous step fiber end face is adsorbed on fiber end face, its other end then carries amino, the immersion time of fiber end face is 24 hours, fiber end face and APTES are reacted fully, covalent bond carry out silane coupledization, and completes amido modified at end face; Biotin (NHS-Biotin) is finally used to modify fiber end face, biotin can in previous step fiber end face modify amino covalence is combined formation stablize conjugate, fiber end face is made to carry biotin molecule like this, define specific adsorption rete, the biotin of rete can produce very strong suction-operated with the Streptavidin molecule in solution to be measured, thus completes the making of optical fiber probe.
System operating mode of the present utility model is: the light signal that light supply apparatus (10) sends enters from three port circulator (40) first ports through Transmission Fibers (30), arrive the second port and arrive through Transmission Fibers (30) and arrive optical fiber probe (60), because interfacial both sides, fiber end face place refractive index is different, Fresnel reflection will be there is, reflected light signal returns along Transmission Fibers (30), arrive the 3rd port through three ports light rings (40) second ports and received by spectroanalysis instrument (20) by Transmission Fibers (30), the intensity of its reflected light is subject to the impact of end face interphase both sides refractive index.Optical fiber probe (60) is vertically placed, and fixed, and end immerses in solution to be measured (70) completely by optical fiber stationary fixture (50), is shifted out by optical fiber probe and drying in atmosphere after 15 minutes from solution.The biotin molecule that fiber end face carries only plays selectivity Specific adsorption to Streptavidin molecule, for the solution of streptavidin due to variable concentrations, the Streptavidin molecular amounts that the biotin molecule of fiber end face absorbs is different, cause the refractive index of fiber end face different, thus the light intensity making spectroanalysis instrument receive is different, the concentration information of Streptavidin converts the intensity of light signal to realize detecting the most at last.
Fresnel reflection occurs in the interphase of two kinds of different refractivity materials, and reflection strength can according to following formulae discovery:
R＝P
r/P
i＝((n
1-n
2)/(n
1+n
2))
2
In formula, R is fiber end face reflectivity, and it equals reflective light intensity (luminous power) P
rwith incident intensity (luminous power) P
iratio; Reflectivity R also can by the refractive index n on end face interphase both sides
1, n
2calculate, wherein n
1for the refractive index of inner fibre core, n
2for end face opposite side is through modifying the refractive index after absorption Streptavidin, i.e. n
2change along with the concentration of solution of streptavidin.At incident optical power P
iwith fiber core refractive index n
1under definite value obtains situation, utilize above-mentioned formula, by measuring reflected optical power P
rsize both can obtain the refractive index n that end face carries Streptavidin side
2, finally can obtain the concentration of solution of streptavidin.
In the present embodiment, the output wavelength of light supply apparatus is 1200nm ~ 1650nm; The operation wavelength coverage of spectroanalysis instrument is 1200nm ~ 1650nm, when detecting luminous power, choose certain wavelength and detect its luminous power, then light source can change single wavelength laser into, spectroanalysis instrument can change conventional light power meter into, can reduce costs; The optical fiber of Transmission Fibers and making optical fiber probe all adopts Standard single-mode fiber (G.652), and core diameter is 8.2 microns, and cladding diameter is 125 microns, and numerical aperture is 0.14.
Disclosed the utility model by above embodiment, but protection domain of the present utility model is not limited thereto, under the condition not departing from the present invention's design, above each component can be replaced with the similar or equivalent element that art personnel understand.
Claims (2)
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201420568366.4U CN204101452U (en) | 2014-09-29 | 2014-09-29 | Based on the Streptavidin concentration sensor of single-mode fiber |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201420568366.4U CN204101452U (en) | 2014-09-29 | 2014-09-29 | Based on the Streptavidin concentration sensor of single-mode fiber |
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| Publication Number | Publication Date |
|---|---|
| CN204101452U true CN204101452U (en) | 2015-01-14 |
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ID=52269871
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112869738A (en) * | 2021-03-17 | 2021-06-01 | 浙江大学 | Optical fiber type real-time cytokine sensing device and preparation method thereof |
| CN113424053A (en) * | 2019-02-08 | 2021-09-21 | 弗朗什孔岱大学 | Method for manufacturing pH measuring sensor based on optical fiber |
-
2014
- 2014-09-29 CN CN201420568366.4U patent/CN204101452U/en not_active Expired - Fee Related
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113424053A (en) * | 2019-02-08 | 2021-09-21 | 弗朗什孔岱大学 | Method for manufacturing pH measuring sensor based on optical fiber |
| CN112869738A (en) * | 2021-03-17 | 2021-06-01 | 浙江大学 | Optical fiber type real-time cytokine sensing device and preparation method thereof |
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| CF01 | Termination of patent right due to non-payment of annual fee |
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