CN100470279C - Biosensor of full fiber optic evanescent wave - Google Patents
Biosensor of full fiber optic evanescent wave Download PDFInfo
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- CN100470279C CN100470279C CNB2006100894974A CN200610089497A CN100470279C CN 100470279 C CN100470279 C CN 100470279C CN B2006100894974 A CNB2006100894974 A CN B2006100894974A CN 200610089497 A CN200610089497 A CN 200610089497A CN 100470279 C CN100470279 C CN 100470279C
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Abstract
The invention relates to a full optical fiber instant passing wave biology sensor technology to realize biological detection by using laser to excite the biology material to emitting. The feature is that the laser transferring and fluorescence receiving path of the sensor contains an optical fiber probe, a single and multiple optical fiber coupler. The laser from the laser emitting device would send to the end of the single-mode optical fiber, and the other end coupling to multimode fiber. One end of the multimode fiber connects to fiber probe, and the other end locating in the light incidence end of fluorescence collecting and data processing device. The invention could decrease the consumption, improve coupling efficiency and decrease optical discrete element and simplify instrument structure.
Description
Technical field:
Biosensor of full fiber optic evanescent wave belongs to technical field of biological, specially refers to that to utilize laser excitation to be marked with the biological substance of fluorescence molecule luminous, thereby realizes the technology of biological detection.
Background technology:
Biologic sensor for fast travelling waves of optical fibre produces evanescent wave when utilizing light wave to transmit in the total reflection mode in the residing medium of fibre-optical probe in optical fiber, this evanescent wave can excite by absorption or with biological compatible reaction and be connected the biological substance that is marked with fluorescence molecule on the detecting head surface, thereby realize detecting the purpose of target substance concentration to be measured.Usually the penetration depth of evanescent wave field has only tens nanometer to the hundreds of nanometer, be incorporated into the fluorescence that the fluorescence molecule in the scope of evanescent wave field sends so biologic sensor for fast travelling waves of optical fibre can only detect, and free fluorescence molecule is contributed to testing result in the solution very little.Therefore biologic sensor for fast travelling waves of optical fibre have highly sensitive, biologic specificity is strong; Simple to operate, measuring speed is fast; Can monitor the dynamic process of biological respinse; Complete machine can miniaturization etc. characteristics, thereby make it obtain using widely in biomedical research, food inspection, environmental monitoring, biological warfare agent field of detecting.
Traditional biologic sensor for fast travelling waves of optical fibre mainly is to utilize optical fiber as probe and optical information hop, and utilizes separating component in a large number in the parts such as collection of signal.Transmission, collection and the detection of the fluorescence that goes out to exciting light with from fibre-optical probe one end-fire all is to adopt conventional optical system, generally includes optical elements such as catoptron, lens combination, chopper.Therefore there is system complex; Energy loss is big; Element positioning requirements height; Big, the difficult instrument miniaturization of realizing takes up room; The light path arrangement flexibility ratio is low, regulates shortcomings such as difficulty is big.As document 1[G.Duveneck, M.Ehrat, H.M.Widmer.Fiber optic evanescentwave biosensor (biologic sensor for fast travelling waves of optical fibre) .SPIE, 1991,1510:138～145]; The yellow favour outstanding person of document 2[, Zhai Junhui, an ice is strong etc., biologic sensor for fast travelling waves of optical fibre and application thereof, optics journal, 2003,23 (4): 451～454].
By retrieval, do not relate to the technical scheme of related content of the present invention in the present disclosed document.
Summary of the invention:
The objective of the invention is to, a kind of biosensor of full fiber optic evanescent wave is proposed, the transmission that the present invention utilizes list-multi-module optical fiber coupler to combine with fibre-optical probe to finish exciting light and the collection and the transmission of fluorescence, single-mode fiber can effectively reduce the energy loss of light transmission, the multimode optical fiber that the fluorescence receiving light path adopts not only can increase the fluorescence quality that detecting head surface can be excited, and can improve the coupling efficiency of fluorescence, so just, the transmission efficiency of light and the signal to noise ratio (S/N ratio) of instrument have been improved, and reduced the optical fractionation element as much as possible, simplified apparatus structure.Adopt the joints of optical fibre can realize the sequence detection of several samples simultaneously.
The biologic sensor for fast travelling waves that the present invention proposes, contain laser beam emitting device, Laser Transmission and fluorescence receiving light path, sample flow path, fluorescence is gathered and data processing equipment, it is characterized in that, described Laser Transmission and fluorescence receiving light path contain a fibre-optical probe (8) and a list-multi-module optical fiber coupler (5) that places sample flow path, described single multi-module optical fiber coupler (5) contains a single-mode fiber (501) and a root multimode fiber (502), the laser emitting end of described laser beam emitting device is coupled to an end of described single-mode fiber (501) to fiber coupler (2) by a laser, the other end of described single-mode fiber (501) is coupled in the described multimode optical fiber (502), one end of described multimode optical fiber (502) connects described fibre-optical probe (8), and its other end places the light incident side of described fluorescence collection and data processing equipment; Described fibre-optical probe (8) is the tapered optical fiber of combination, and this fibre-optical probe (8) finishing has biological identification molecule.
Described multimode optical fiber (502) can be connected with described fibre-optical probe (8) by joints of optical fibre (6).
Evidence, biology sensor proposed by the invention can effectively reduce the energy loss of light transmission, improves the coupling efficiency of fluorescence, has further improved the transmission efficiency of light and the signal to noise ratio (S/N ratio) of instrument, and reduced the optical fractionation element as much as possible, simplified apparatus structure.
Description of drawings:
Fig. 1 is the structural representation of biosensor of full fiber optic evanescent wave of the present invention.
Fig. 2 is the list-multi-module optical fiber coupler structural representation in the biosensor of full fiber optic evanescent wave of the present invention.
Fig. 3 is the fibre-optical probe structural representation in the biosensor of full fiber optic evanescent wave of the present invention.
Embodiment:
See also Fig. 1, Fig. 1 is the structural representation of biosensor of full fiber optic evanescent wave of the present invention.As seen from the figure, biosensor of full fiber optic evanescent wave of the present invention comprises laser beam emitting device, Laser Transmission and fluorescence receiving light path, sample flow path, fluorescence harvester and data processing equipment.Wherein laser beam emitting device contains pulsed laser 1 and pulse signal generator 10; Laser Transmission and fluorescence receiving light path contain laser to fiber coupler 2, list-multi-module optical fiber coupler 5 and fibre-optical probe 8; Sample flow path contains sample cell 9, peristaltic pump 13, and sample cell inlet 7 and sample cell outlet 14 are arranged on the sample cell; Fluorescence collection and data processing equipment contain fluorescent optical filter 4, photodetector 3, lock-in amplifier 11 and computing machine 12.
As seen from Figure 1, the laser that pulsed laser 1 sends enters single-mode fiber 501 in list-multi-module optical fiber coupler 5 by laser to fiber coupler 2, multimode optical fiber 502 transmission through list-multi-module optical fiber coupler 5 enter fibre-optical probe 8 then, produce evanescent wave in fibre-optical probe 8 near surfaces zones, be connected the fibre-optical probe 8 lip-deep biological substances that are marked with fluorescence molecule and send fluorescence to excite.Part fluorescence coupled back into optical fibers pops one's head in 8, enters the multimode optical fiber 502 of list-multi-module optical fiber coupler 5, in the other end ejaculation of multimode optical fiber 502.Fluorescent optical filter 4 filterings exciting light reflection or that lose, and most of fluorescence is seen through, the fluorescence that sees through is surveyed by photodetector 3 (can adopt photodiode) and is converted light signal to electric signal, this electric signal amplifies through lock-in amplifier 11 then, gathers the line data analysis of going forward side by side by computing machine 12.The pulse signal of pulsed laser 1 is provided by pulse signal generator 10, and pulse signal generator 10 provides the same frequency signal as the reference signal for lock-in amplifier 11 simultaneously.
As shown in Figure 2, principal feature of the present invention is to have adopted list-multi-module optical fiber coupler 5 to substitute Laser Transmission and fluorescence receiving light path that optical elements such as traditional reflective mirror, lens combination are formed.List-the multi-module optical fiber coupler that adopts is to draw the awl method to make with welding commonly used at present, wherein single-mode fiber 501 is introduced multimode optical fiber as the Laser Transmission light path with exciting light, because single-mode fiber only transmits a kind of light of pattern, thereby can stop the loss of light effectively.And the fluorescence receiving light path adopts multimode optical fiber, not only can increase the fluorescence quality that detecting head surface can be excited, and can improve the coupling efficiency of fluorescence, thus the sensitivity and the signal to noise ratio (S/N ratio) of raising system.
As shown in Figure 3, fibre-optical probe 8 adopts the conventional tapered form of combination, and the part covering 801 of optical fiber is removed, and making up tapered probe 802 finishinges has biological identification molecule.Like this, can improve the incident angle of incident light on the one hand, make incident light increase, increase the number of times that excites, thereby can inspire more fluorescent energy fluorescence molecule at fibre-optical probe 8 internal reflection number of times; On the other hand, the fluorescence that is coupled into probe with the high-order mode form can be changed into low-order mode, reduce the fluorescence losses that does not match and cause because of pattern, with collection and the transfer efficiency of further raising system fluorescence.
Fibre-optical probe 8 can be an end of multimode optical fiber, but being the sequence detection of sample, this structure makes troubles, for the sequence detection several samples, can adopt dismountable joints of optical fibre 6 (as FC, SC, ST type etc.) that fibre-optical probe 8 is connected with single multi-module optical fiber coupler 5.After using the joints of optical fibre 6, after having detected a certain sample, can be replaced with the probe that another is modified with other biological identification molecule, to realize the detection of other sample.
Identical with the using method of traditional biosensor of full fiber optic evanescent wave, before biosensor of full fiber optic evanescent wave of the present invention detects, need to modify biological identification molecule at probe 8, will pop one's head in then 8 places sample cell 9.Open peristaltic pump 13, testing sample is sucked sample cell 9, behind the reaction certain hour, open pulsed laser 1, exciting light will enter probe 8 by list-multi-module optical fiber coupler 5 in the total reflection mode, the evanescent wave that laser beam produces during multihop propagation in probe 8 excites the fluorescence molecule of detecting head surface, fluorescence molecule sends fluorescence, part is coupled back and pops one's head in 8, after multimode optical fiber 502 transmission, penetrate from multimode optical fiber 502 other ends, through the fluorescent optical filter 4 laggard photodetector 3 of going into that filters, be converted into the electric signal that is directly proportional with fluorescence signal intensity, amplify the back through lock-in amplifier 11 and gathered by computing machine 12.The electric signal that computing machine 12 collects is directly proportional with the concentration of detecting head surface measured matter, by the amplitude of analyzing electric signal can obtain the popping one's head in concentration of 8 surperficial measured matters.
The present invention has provided an embodiment, and its concrete structure and parameter are as follows:
LASER Light Source is that centre wavelength is the semiconductor laser 1 of 635nm, and output power is 8mW, and laser is to coupling efficiency 〉=20% of fiber coupler 2.The optical fiber of list-multi-module optical fiber coupler 5 is silica fibre, and wherein the diameter of single-mode fiber is 3 μ m, and numerical aperture is 0.11; The diameter of multimode optical fiber 502 is 600 μ m, and numerical aperture is 0.22.Transmitance≤10 of 4 pairs of exciting lights of fluorescent optical filter
-7, to transmitance 〉=80% of fluorescence.Photodetector 3 is a photodiode.
During measurement, the biological sample solution of Cy5.5 fluorescence molecule that in sample cell 6, injected mark, every fibre-optical probe all reaches 10 to the detection sensitivity of Cy5.5 fluorescence molecule solution among the embodiment
-10Mol/L.The detection sensitivity difference of each fibre-optical probe is less than 5%.Finish first order fluorescence input required time and be less than 15min.
Claims (2)
1, biosensor of full fiber optic evanescent wave, contain laser beam emitting device, Laser Transmission and fluorescence receiving light path, sample flow path, fluorescence is gathered and data processing equipment, it is characterized in that, described Laser Transmission and fluorescence receiving light path contain a fibre-optical probe (8) and a single multi-module optical fiber coupler (5) that places sample flow path, described single multi-module optical fiber coupler (5) contains a single-mode fiber (501) and a root multimode fiber (502), the laser emitting end of described laser beam emitting device is coupled to an end of described single-mode fiber (501) to fiber coupler (2) by a laser, the other end of described single-mode fiber (501) is coupled in the described multimode optical fiber (502), one end of described multimode optical fiber (502) connects described fibre-optical probe (8), and its other end places the light incident side of described fluorescence collection and data processing equipment; Described fibre-optical probe (8) is the tapered optical fiber of combination, and this fibre-optical probe (8) finishing has biological identification molecule.
2, biosensor of full fiber optic evanescent wave according to claim 1 is characterized in that, described multimode optical fiber (502) is connected with described fibre-optical probe (8) by joints of optical fibre (6).
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101545899B (en) * | 2009-04-30 | 2012-12-05 | 华南理工大学 | Optical fibre micro-fluidic biological sensor and preparation method thereof |
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| CN101008649B (en) * | 2007-01-26 | 2011-04-06 | 中国科学院上海光学精密机械研究所 | Multi-probe optical fiber evanescent wave biological detector based on correlation detection |
| CN101701913B (en) * | 2009-11-20 | 2011-03-30 | 清华大学 | Multi-probe optical fiber evanescent wave biological sensor with all-optical-fiber structure |
| CN101949849B (en) * | 2010-09-08 | 2011-09-21 | 华中科技大学 | Light-activated positioning microscopic imaging system based on optical fiber evanescent field illuminator |
| CN103245641B (en) * | 2012-02-10 | 2014-10-29 | 清华大学 | Multi-channel planar waveguide evanescent wave biosensor |
| CN103056513B (en) * | 2012-12-14 | 2014-12-10 | 武汉锐科光纤激光器技术有限责任公司 | Laser processing system |
| CN103335992B (en) * | 2013-06-21 | 2016-02-10 | 北京交通大学 | A kind of fluorescence type glucose capillary biosensor |
| CN103486975B (en) * | 2013-10-16 | 2015-12-23 | 重庆大学 | Biofilm thickness optical evanescent wave sensor device and Biosensor for Detecting Bio-layer system thereof |
| CN106198464A (en) * | 2015-05-08 | 2016-12-07 | 罗强 | A kind of laser-induced fluorescence (LIF) food detector |
| CN105115955A (en) * | 2015-09-16 | 2015-12-02 | 北京首量科技有限公司 | Optical fiber detection device for biological detection |
| CN105424663A (en) * | 2015-11-24 | 2016-03-23 | 西南大学 | Method for detecting phthalic acid ester compound concentration based on optical fiber immunosense |
| CN105784654A (en) * | 2016-03-11 | 2016-07-20 | 清华大学 | Optical fiber fluorescent sensor capable of detecting varied microorganisms simultaneously |
| CN105606583A (en) * | 2016-03-25 | 2016-05-25 | 北京首量科技有限公司 | Optical fiber detection system for biological detection |
| CN106198471B (en) * | 2016-05-06 | 2019-04-09 | 黄辉 | A kind of bio-chemical fluorescent analyzer and its detection method based on light-conducting capillaries |
| CN106990086B (en) * | 2017-06-06 | 2020-02-28 | 中国人民大学 | Multi-channel evanescent wave all-fiber biosensor |
| CN107988319B (en) * | 2017-12-13 | 2021-04-09 | 清华大学 | Method for evaluating interaction of two single-stranded nucleic acid molecules |
| CN112083159B (en) * | 2019-06-13 | 2022-03-25 | 首都师范大学 | Evanescent wave aptamer sensor and method for detecting small molecules by applying evanescent wave aptamer sensor |
| CN110243795B (en) * | 2019-06-19 | 2022-12-02 | 中国人民大学 | Evanescent wave bicolor fluorescent all-fiber biosensor |
| CN113237859A (en) * | 2021-05-11 | 2021-08-10 | 国网安徽省电力有限公司电力科学研究院 | Diamond film fluorescence high-efficiency collecting device |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101545899B (en) * | 2009-04-30 | 2012-12-05 | 华南理工大学 | Optical fibre micro-fluidic biological sensor and preparation method thereof |
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Assignee: Beijing Jinda Qingchuang Environmental Science and Technology Co., Ltd. Assignor: Tsinghua University Contract record no.: 2011990000909 Denomination of invention: Biosensor of full fiber optic evanescent wave Granted publication date: 20090318 License type: Exclusive License Open date: 20061206 Record date: 20110919 |
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