CN102507520B - Liquid core waveguide fluorescence detector - Google Patents
Liquid core waveguide fluorescence detector Download PDFInfo
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- CN102507520B CN102507520B CN 201110343868 CN201110343868A CN102507520B CN 102507520 B CN102507520 B CN 102507520B CN 201110343868 CN201110343868 CN 201110343868 CN 201110343868 A CN201110343868 A CN 201110343868A CN 102507520 B CN102507520 B CN 102507520B
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Abstract
The invention relates to a liquid core waveguide detector, in particular to a liquid core waveguide fluorescence detector, which comprises a liquid core optical fiber, an exciting light source, and a fluorescence detection component. The side wall of the liquid core optical fiber is provided with an incident window; exciting liquid emitted by the exciting light source is emitted into the liquid core optical fiber at a proper angle through the incident window on the side all of the liquid core optical fiber, and is spread in a total reflection way along the axis direction of the liquid core optical fiber; a test sample in a liquid medium in the liquid core optical fiber is excited and emits fluorescence; and the fluorescence detection component detects the fluorescence transmitted through the side wall of the liquid core optical fiber. The liquid core waveguide detector has the advantages that: the exciting light and the fluorescence are separated by a liquid core waveguide principle, and the background noise of the detector can be reduced effectively; the exciting light is spread in a capillary tube along the axis, the exciting optical path and utilization rate are greatly increased, and the sensitivity of the detector is improved; meanwhile, the device is simple in structure, convenient to use, low in cost and wide in application range.
Description
Technical field
The present invention relates to a kind of liquid core waveguide pick-up unit, specifically a kind of liquid core waveguide fluorescence testing apparatus.
Background technology
(Liquid Core Waveguide, LCW) phenomenon is called liquid-core optical fibre again to liquid core waveguide.Refer to that the refractive index when perimeter material is lower than the refractive index of liquid core, and the incident angle of incident ray is during greater than critical angle of incidence, total reflection takes place at the interface of liquid and duct wall in light, light in certain angle will can almost can't harm the lost territory from the phenomenon of other end conduction output along the axial propagation of liquid core.
The research of LCW starts from the middle of the 19th century the earliest, and people such as Colladon, Babinet, Tyndall have studied the phenomenon that light is propagated in stream by total internal reflection.To 19 end of the centurys, this phenomenon is widely known by the people because of the application in the illumination fountain.About 1970, the researcher begins to note that LCW is used as the possibility of communication media, has made by thin footpath glass tube to be full of the LCW that high refractive index liquid constitutes.But because solid optical fiber has remarkable advantages, make that LCW fails to be applied in communication.But it has but represented tempting application prospect on analytical chemistry, has been applied to various optical detection such as long light path absorption detecting, chemiluminescence detection, fluoroscopic examination, Raman spectrum detection system.
Robert H.Byrne is used for photometer with liquid core waveguide, and CCD when the uviol lamp of instrument the inside is closed, as ultraviolet-visible spectrophotometer, when uviol lamp is opened, can be used as fluorophotometer as detecting device; Vratislav Kostal sets up fluorescence detecting system with liquid core waveguide and optical fiber coupled CCD photometer, and test with the protein of low-molecular-weight fluorescent marker and fluorescein isothiocyanate (FITC) mark, comparing with the laser induced fluorescence detector of complexity not only has better detectability, and it is as long as the repacking light source just can be analyzed other analytic targets, thereby more convenient.
Johan Roeraade etc. are with LCW and Capillary Electrophoresis laser-induced fluorescence (LIF) coupling CCD end point detection, under continuous flow and capillary electrophoretic, can detect 2.7pmol/L and 62fmol/L fluorescein respectively, they also with the development detector applies in dna sequence analysis, differentiate monochromatic G sequence base, when signal to noise ratio (S/N ratio) is 250,500 bases can be detected, when signal to noise ratio (S/N ratio) is 50,950 bases can be detected.The imaging of report such as Liu Zhen capillary isoelectric focusing liquid core waveguide electrophoresis laser-induced fluorescence (LIF) integral post detects, and detects with business-like ultraviolet and compares, and sensitivity can improve 3-5 the order of magnitude.This type of liquid Core Waveguide Technique exciting light and fluorescence signal spread out of jointly from an end, collect and detect, and fail to realize preferably separating of exciting light and fluorescence, and there is bigger interference in testing result.Therefore inventing the liquid core waveguide pick-up unit that can realize better that exciting light separates with fluorescence has great importance.
Summary of the invention
The objective of the invention is to: a kind of liquid core waveguide fluorescence testing apparatus is provided, this device is used the liquid core waveguide principle, exciting light is injected from the kapillary sidewall with suitable angle, and in coat inwall generation total reflection, and propagate along the kapillary axis, fluorescence will be gone out from the capillary tube inner wall transmission simultaneously, enter photoelectric detector, and realization fluorescence separates with exciting light.
The objective of the invention is to be achieved through the following technical solutions: a kind of liquid core waveguide fluorescence testing apparatus, comprise liquid-core optical fibre, exciting light sources and fluoroscopic examination assembly, the sidewall of liquid-core optical fibre has incidence window, the exciting light that exciting light sources sends is injected liquid-core optical fibre with a suitable angle from the incidence window of liquid-core optical fibre sidewall, in liquid-core optical fibre, propagate along the axis direction total reflection, after being excited, specimen in the liquid medium in the liquid-core optical fibre launches fluorescence, the fluorescence that the fluoroscopic examination component detection transmits from the sidewall of liquid-core optical fibre.
The fluoroscopic examination assembly comprises fluorescence gathering system, filter system and photoelectric detector, is collected by fluorescence gathering system from the fluorescence that the sidewall of liquid-core optical fibre transmits, and is received by photoelectric detector behind filter system.
Liquid-core optical fibre is by constituting for coat, quartz layer and the liquid medium that optical index is increased successively from outside to inside.
Exciting light sources is laser instrument, light emitting diode, deuterium lamp or xenon lamp, between the incidence window of exciting light sources and liquid-core optical fibre diaphragm is set.
Fluorescence gathering system comprises condenser lens and concave mirror, lay respectively at the both sides of liquid-core optical fibre, photoelectric detector is positioned at condenser lens one side, filter system is between condenser lens and photoelectric detector, the fluorescence that points to photoelectric detector directly is focused lens and focuses on, and rightabout fluorescence is focused on by condenser lens by the concave reflection mirror reflection, then by being received by photoelectric detector behind the filter system removal veiling glare.
Filter system is made up of optical filter and diaphragm.
Photoelectric detector is photomultiplier or CCD.
The present invention has the following advantages:
1, total reflection takes place in liquid-core optical fibre inside in exciting light among the present invention, and propagates along axis direction, has farthest increased exciting light light path and utilization factor, thereby has improved the sensitivity of detecting device greatly.
2, realized separating of exciting light and fluorescence, eliminated the interference of exciting light to testing result, reduced ground unrest, improved the detecting device signal to noise ratio (S/N ratio).
3, the concave mirror fluorescence that will deviate from photoelectric detector reflexes to the photoelectric detector direction, has increased phosphor collection efficient, has improved detection sensitivity.
4, liquid core waveguide fluorescence testing apparatus is simple in structure, and is easy to use, with low cost, and usable range is wide.
Description of drawings
Fig. 1 is liquid core waveguide fluorescence testing apparatus synoptic diagram of the present invention.
Fig. 2 is that exciting light of the present invention and fluorescence signal are propagated the light path synoptic diagram in quartz capillary.
Fig. 3 is the analysis chromatogram of amino acid mixing sample among the embodiment 1.
Mark in the accompanying drawing is respectively: 1. exciting light sources, 2. exciting light light, 3. fluorescence, 4. condenser lens, 5. photoelectric detector, 6. optical filter, 7. shadow shield, 8. liquid-core optical fibre, 9. concave mirror, 10. coat, 11. quartz layers, 12. liquid mediums.
Embodiment
A kind of liquid core waveguide fluorescence testing apparatus comprises liquid-core optical fibre 8, exciting light sources 1 and fluoroscopic examination assembly, and exciting light sources 1 and fluoroscopic examination assembly are positioned at the side of liquid-core optical fibre 8.The fluoroscopic examination assembly comprises fluorescence gathering system, filter system and photoelectric detector 5, liquid-core optical fibre 8 passes fluorescence gathering system, filter system is between photoelectric detector 5 and fluorescence gathering system, the fluorescence that transmits from the sidewall of liquid-core optical fibre 83 is collected by fluorescence gathering system, is received by photoelectric detector 5 behind filter system.
Wherein liquid-core optical fibre 8 comprises the outer refraction coefficient that applies of quartz capillary, quartz capillary less than the Teflon AF coating material of the quartz coat 10 as quartz capillary, and at one end removes a small pieces coating material, as incidence window; Quartz capillary inside be refraction coefficient greater than the liquid medium 12 of quartz, whole liquid-core optical fibre 8 is the close-structure of dredging for refraction index, namely the coat 10 at quartz capillary forms the interior wall construction that one decks have the total reflection ability.
Wherein fluorescence gathering system comprises condenser lens 4 and concave mirror 9, lays respectively at both sides capillaceous.Filter system is made up of diaphragm and optical filter 6, between photoelectric detector 5 and condenser lens 4, fluorescence 3 is focused lens 4 after the mating plate 6 after filtration and focuses on the diaphragm center, and diaphragm and optical filter 6 actings in conjunction make that the light signal that enters in the photoelectric detector 5 is pure fluorescence 3.
In liquid core waveguide fluorescence testing apparatus, the exciting light that exciting light sources 1 sends is injected from liquid-core optical fibre 8 sides, 10 total reflection of coated layer, and in liquid-core optical fibre 8 inside along its axis propagation, and the fluorescence 3 of specimen emission can transmit from liquid-core optical fibre 8 sidewalls, collected by fluorescence gathering system and filter by filter system that veiling glare is laggard goes into photoelectric detector 5, realize separating of exciting light and fluorescence 3.
Be specially: the inner filled with liquid medium 12 of quartz capillary, total is liquid medium 12, quartz layer 11 and Teflon AF coat 10 from the inside to surface, refraction coefficient reduces successively.After exciting light enters liquid-core optical fibre 8 from incidence window at a certain angle, in quartz layer 11, reflect, to go into to inject at a certain angle in the liquid medium 12, the tested sample of a part of light absorbs, and inspires fluorescence 3 and consumes.According to the light path principle of reversibility, the exciting light of Xiao Haoing is not propagated in the relative quartz layer 11 of incidence window with equal angular.Select suitable incident angle, make the angle of direction that exciting light propagates at quartz layer 11 and coat 10 normals greater than the cirtical angle of total reflection of exciting light at coat 10, thereby in coated layer of 10 reflected back liquid medium 12.So repeat, exciting light is propagated in liquid-core optical fibre 8, until its endpiece, and can not penetrate from the side.Specimen is excited and launches fluorescence 3 in the liquid medium 12, this fluorescence 3 can be regarded pointolite as, it is transmitted to coat 10 and angle and penetrates less than the fluorescence 3 of the cirtical angle of total reflection sidewall from liquid-core optical fibre 8, collected by fluorescence gathering system then and enter photoelectric detector 5, thereby realize separating of fluorescence 3 and exciting light.The exciting light of propagating along liquid-core optical fibre 8 does not stop to excite specimen in whole liquid-core optical fibre 8 simultaneously, specimen does not stop to be excited in liquid-core optical fibre 8, launches fluorescence 3, has improved the utilization factor of exciting light greatly, increase available fluorescence 3, and then improved detection sensitivity.
As shown in Figure 1, liquid-core optical fibre 8 employing specifications are the quartz capillary that 50 μ m ID * 45/70cm (separation length/length overall) Teflon AF2400 apply.Exciting light sources 1 is laser instrument, and liquid-core optical fibre 8 is made up of coat 10, quartz layer 11, liquid medium 12 3 parts, and its refraction coefficient increases successively.Shadow shield 7 is the black plastic sheet of internal diameter 0.4mm thickness 0.5mm, and this shadow shield 7 is the diaphragm in the filter system, and optical filter 6 is the logical optical interference filter of the length of cutoff wavelength 520nm.Liquid-core optical fibre 8 one ends remove a small pieces coat 10 as incidence window, and the exciting light light 2 of exciting light sources 1 emission enters liquid-core optical fibre 8 inside with certain angle from incidence window.Total reflection takes place at coat 10 inwalls in exciting light light 2, coated layer 10 reflected back liquid-core optical fibre 8 inside, and along axis propagation, constantly exciting liquid body medium 12 produces fluorescence 3 simultaneously.Fluorescence 3 will be gone out from liquid-core optical fibre 8 sidewall transmissions, deviate from a part of fluorescence 3 of photoelectric detector 5 through concave mirror 9 reflections, focus on through condenser lens 4 with opposite side fluorescence 3 and collect, and optical filter 6 is removed veiling glare, enters in the photoelectric detector 5.
Fig. 3 analyzes the detection collection of illustrative plates of fluorescein isothiocynate (FITC) derivatization arginine, leucine, glycocoll recombined sample for the liquid core waveguide fluorescence testing apparatus among the use embodiment 1.In the mixed amino acid derivative sample concentration of FITC-arginine derivative, FITC-leucine derivant and FITC-glycine derivative be respectively 0.1,0.2,0.1nmol/L, use the laser instrument of centre wavelength 473nm, power 10mW as exciting light sources 1.The electrophoretic separation condition: back-ground electolyte is the 80mmol/L borax soln (pH is 9.2) that contains the 1.0mmol/L hexane diamine; Adopt the mode of pressure sample introduction: 0.5psi (3.45KPa)) * 5s; 22 ℃ of separation voltage 20kV, column temperatures; Arginine detects and is limited to 3.0pmol/L (S/N=3).
Claims (5)
1. liquid core waveguide fluorescence testing apparatus, it is characterized in that: comprise liquid-core optical fibre (8), exciting light sources (1) and fluoroscopic examination assembly, the sidewall of liquid-core optical fibre (8) has incidence window, the exciting light that exciting light sources (1) sends is injected liquid-core optical fibre (8) with a suitable angle from the incidence window of liquid-core optical fibre (8) sidewall, in liquid-core optical fibre (8), propagate along the axis direction total reflection, after being excited, specimen in the liquid medium (12) in the liquid-core optical fibre (8) launches fluorescence (3), the fluorescence (3) that the fluoroscopic examination component detection transmits from the sidewall of liquid-core optical fibre (8);
Described fluoroscopic examination assembly comprises fluorescence gathering system, filter system and photoelectric detector (5), is collected by fluorescence gathering system from the fluorescence (3) that the sidewall of liquid-core optical fibre (8) transmits, and is received by photoelectric detector (5) behind filter system;
Liquid-core optical fibre (8) passes fluorescence gathering system, described fluorescence gathering system comprises condenser lens (4) and concave mirror (9), lay respectively at the both sides of liquid-core optical fibre (8), photoelectric detector (5) is positioned at condenser lens (4) one sides, filter system is positioned between condenser lens (4) and the photoelectric detector (5), the fluorescence (3) that points to photoelectric detector (5) directly is focused lens (4) and focuses on, and rightabout fluorescence (3) is focused on by condenser lens (4) by concave mirror (9) reflection, then by being received by photoelectric detector (5) behind the filter system removal veiling glare.
2. liquid core waveguide fluorescence testing apparatus according to claim 1 is characterized in that: described liquid-core optical fibre (8) is by coat (10), quartz layer (11) and liquid medium (12) formation for optical index is increased successively from outside to inside.
3. liquid core waveguide fluorescence testing apparatus according to claim 1, it is characterized in that: described exciting light sources (1) arranges diaphragm for laser instrument, light emitting diode, deuterium lamp or xenon lamp between the incidence window of exciting light sources (1) and liquid-core optical fibre (8).
4. liquid core waveguide fluorescence testing apparatus according to claim 1, it is characterized in that: described filter system is made up of optical filter (6) and diaphragm.
5. liquid core waveguide fluorescence testing apparatus according to claim 1, it is characterized in that: described photoelectric detector (5) is photomultiplier or CCD.
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| CN103063645B (en) * | 2013-01-04 | 2017-12-05 | 南开大学 | High-efficiency fluorescence detection based on novel Microstructure optical fiber |
| CN103105373A (en) * | 2013-01-14 | 2013-05-15 | 中国农业大学 | Dynamic spectrophotometry testing system and method for benzene in water |
| CN106353294A (en) * | 2016-11-09 | 2017-01-25 | 武汉能斯特科技有限公司 | Miniature capillary luminoscope |
| CN107643273B (en) * | 2017-10-20 | 2023-09-08 | 中国计量大学 | Aggregation-induced emission biosensor based on hollow photonic crystal fiber |
| EP3492909B1 (en) * | 2017-12-01 | 2023-11-01 | ams AG | Chemical sensing device using fluorescent sensing material |
| CN110260256A (en) * | 2019-05-29 | 2019-09-20 | 深圳市星汉激光科技有限公司 | A kind of white light laser source |
| CN112485734B (en) * | 2020-09-27 | 2022-12-13 | 中国电子科技集团公司第十三研究所 | Method for improving fluorescence collection efficiency of NV color centers of diamonds |
| CN113155795B (en) * | 2021-04-15 | 2023-03-21 | 西北核技术研究所 | Device and method for directly measuring upper energy level fluorescence lifetime of rare earth element doped optical fiber laser |
| CN113109314A (en) * | 2021-05-28 | 2021-07-13 | 上海睿钰生物科技有限公司 | Multiple fluorescence signal detection system and method |
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| WO2009088492A1 (en) * | 2008-01-04 | 2009-07-16 | The Government Of The United States Of America, As Represented By The Secretary Of The Navy Naval | Optical separator and method for separating particles suspended in a fluid |
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| US7211763B2 (en) * | 2004-12-22 | 2007-05-01 | General Electric Company | Photon energy material processing using liquid core waveguide and a computer program for controlling the same |
| US20110149286A1 (en) * | 2009-12-18 | 2011-06-23 | Chih-Wei Wu | Liquid core waveguide assembly and detecting system including the same |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| WO2009088492A1 (en) * | 2008-01-04 | 2009-07-16 | The Government Of The United States Of America, As Represented By The Secretary Of The Navy Naval | Optical separator and method for separating particles suspended in a fluid |
| CN101271070A (en) * | 2008-05-09 | 2008-09-24 | 东北大学 | Microcurrent controlled capillary tube electrophoresis liquid core waveguide fluorescence testing apparatus |
| CN101806726A (en) * | 2010-03-29 | 2010-08-18 | 浙江大学 | Double-wavelength absorbance detection device for analyzing trace liquid |
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