CN1167945C - Laser induced fluorescence detector - Google Patents
Laser induced fluorescence detector Download PDFInfo
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- CN1167945C CN1167945C CNB011238623A CN01123862A CN1167945C CN 1167945 C CN1167945 C CN 1167945C CN B011238623 A CNB011238623 A CN B011238623A CN 01123862 A CN01123862 A CN 01123862A CN 1167945 C CN1167945 C CN 1167945C
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- sample cell
- diaphragm
- light source
- induced fluorescence
- fluorescence detector
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Abstract
The utility model relates to a laser induced fluorescence detector which comprises a fine adjusting device, a secondary light source, an observation mirror, an excitation light source, a set of optical lens and a photoelectric signal converter, wherein the fine adjusting device is used for placing and adjusting the position of a sample cell; the secondary light source is used for adjusting a light path; the observation mirror is used for adjusting the light path; the excitation light source is a working light source of the laser induced fluorescence detector and is composed of a laser, reference photoelectric diodes, a weakener and a dichroscope; the set of optical lens is used for aggregating the imaging of the work light source and is composed of a focusing lens, an imaging lens, a light filter and a diaphragm; the photoelectric signal converter is used for receiving and outputting detecting signals. On the basis of a common focus method, the present invention obtains a high efficient imaging system. The changeable sample cell enables the laser induced fluorescence detector to be used for segregation analysis technology, such as micro high-efficiency liquid phase chromatography (Micro-HPLC), capillary electrophoresis (CE), capillary electro chromatography (CEC), etc. The adjustable diaphragm can be used for suppressing stray light farthest, which enables background signals to greatly reduce. A viewing calibrating device enables daily calibration to easily operate.
Description
Technical field
The present invention relates to a kind of laser induced fluorescence detector.
Background technology
The Micro-Column Separation analytic system have traditional compartment analysis system incomparable advantage: post is imitated high, can reduce the consumption of solvent and sample greatly, and environmental pollution is little, is easy to and other detection method on-line coupling.The advantage that the micro pole analysis system is had makes it in environmental science, natural products, and technical fields such as petrochemical complex and biology are extensively utilized.
In micro pole analysis, because sample size very little (receive upgrading), sample concentration is very low again, and this has just proposed very high requirement to detecting device.Laser-Induced Fluorescence Detection has very high sensitivity (ppb-ppt level), laser beam can be converged to a minimum hot spot in addition, be complementary with the microtrabeculae system well, these advantages make laser-Induced Fluorescence Detection become field extremely active in the micro pole analysis system just.
Roughly laser-Induced Fluorescence Detection can be divided into confocal point according to detecting element with the position relation of launching light beam detects and vertical detection.
Confocal point detects owing to simple in structure, phosphor collection rate advantages of higher receive much concern.The subject matter that this pick-up unit exists is difficulty of light path orientation ratio, the output signal of detecting device is very responsive to the position of detection window, it is fixing to add that the kapillary pond is difficult for, this just is difficult to guarantee that the capillary detection pond is in the optimum position all the time, even harmonized light path, also be difficult to guarantee in continued operation, keep the constant of its position, thereby cause analysis result repeatability bad owing to changing the kapillary pond.At present in some commercial confocal laser induced fluorescence detectors, the influence of adopting filter set to deduct exciting light, these filter set are normally combined by several interference filters, sometimes go back breach optical filter of superposition, the interference that exciting light can be effectively deducted in this optical filter combination, but its cost is very high.In addition, the sample cell micromatic setting that in commercial instrument, is equipped with, its mobile accuracy is very high, but size is bigger, is not suitable for being used in the miniature instrument, and the price comparison costliness.
J.Dovichi et.al J.Chromatogr.1989,480:141 has reported in vertical detection according to reversibility of optical path, place a small light source in the photomultiplier front, observe with an attachment lens on the collecting lens opposite, adjust sample cell and condenser lens position, the three is overlapped, reach the purpose of calibration.Desire to reach this purpose, needs one colour glass filter are efficiently deducted exciting light, but the difficult selection of this optical filter if the optical filter efficient of selecting is too high, does not then observe laser spots, if efficient is too low, does not then have due effect.In addition, this optical filter does not have selectivity to exciting light and fill-in light, the deduction exciting light in, be easy to fill-in light is filtered, be difficult to like this observe sample cell clearly as.Anders P.larson et.al Applied Optics, 1993,32 (6): 794 pictures that in finder, utilize an attachment lens under excitation light irradiation, to produce at Direct observation sample cell on the photomultiplier position.The problem that this device exists is, photomultiplier need be removed during observation, and this is very inconvenience for valuable and frangible photomultiplier; Laser shines directly into sample cell can produce a large amount of scattered lights, is difficult to observe directly the accurate position of laser spots, and the easy damaged eyes.
Summary of the invention
The object of the present invention is to provide a kind of laser induced fluorescence detector, can more easily calibrate light path, and the replacing sample cell can not influence calibrated light path in continued operation yet.
A kind of portable laser induced fluorescent tester provided by the invention includes:
One energizer is the work light of detecting device, induces the fluorescer of sample cell to produce fluorescence, is gathered in the photoelectric signal converter output detection signal through the lens combination imaging, and principal feature of the present invention is:
1, sample cell is to be placed on replaceably on the micromatic setting, this micromatic setting middle part is provided with the circular hole of a placement sample cell, moving horizontally bar by adjustment can make cross sliding clock and vertical slipper slide at slideway base, make sample cell do moving up and down within the specific limits, adjust and screw the fixed knob of micromatic setting with the sample cell locking position after finishing;
2, a secondary light source that is used to adjust light path is set between sample cell and the condenser lens;
3, sight glass is screwed into the situation that observation optical path is adjusted on the observation station between diaphragm and the optical filter;
4, adopt the stack of macropore diaphragm and aperture to suppress parasitic light.
The present invention is based on confocal point mode, obtain imaging system efficiently, removable sample cell and micromatic setting make this detecting device can be used on micro high efficiency liquid phase chromatography (Micro-HPLC), and Capillary Electrophoresis (CE) is in the capillary electric chromatogram method compartment analysis technology such as (CEC); Adjustable diaphragm can suppress parasitic light to greatest extent, and background signal is reduced greatly; The observation calibrating installation makes daily calibration be very easy to operation.
Description of drawings
For further understanding the present invention, provide preferred forms below and also be described in detail in conjunction with the accompanying drawings, wherein:
Fig. 1 is an optical system structure synoptic diagram of the present invention;
Fig. 2 a, Fig. 2 b are for passing through the imaging of the observed sample cell of sight glass in the calibration process of the present invention;
Fig. 3 a is the micromatic setting front view of sample cell of the present invention;
Fig. 3 b is the micromatic setting vertical view of sample cell of the present invention;
Fig. 4 a is a macropore diaphragm synoptic diagram of the present invention;
Fig. 4 b is an aperture synoptic diagram of the present invention;
Fig. 5 is the methylene blue chromatogram of 5ppb for concentration of the present invention.
Embodiment
At first please referring to Fig. 1, Fig. 2 a, b and Fig. 3 a, b.
One sample cell 1 places the front end of a lens combination, and this lens combination is made up of condenser lens 3 and imaging len 8.
During the calibration light path, open the secondary light source 2 that is arranged between sample cell 1 and the condenser lens 3, sight glass 9 is screwed on the observation station between diaphragm 11 and the optical filter 10 (being arranged in light path just), wherein diaphragm 11 places the lens combination rear side, optical filter 10 places between diaphragm 11 and the photomultiplier 12, and sample cell 1 is positioned on the micromatic setting 20.The structure of micromatic setting 20 is shown in Fig. 3 a to Fig. 3 b, micromatic setting 20 middle parts are provided with a placement sample cell 1 and are the position 22 of circular hole, and be provided with and move horizontally bar 21, move horizontally bar 21 by adjustment cross sliding clock 23 and vertical slipper 24 are slided at slideway base 25, make sample cell 1 do up and down mobile within the specific limits.
Adjust micromatic setting 20 and lens 3 and 8 make sample cell 1 present at diaphragm 11 places clearly as, Fig. 2 a is laser instrument sample cell 1 imaging in sight glass 9 when not starting, wherein ellipse is the mirror surface in the sight glass 9, four inside and outside wall imagings that vertical line is a sample cell 1.Open then and be arranged on laser instrument 4 light paths and the weakener 5 between condenser lens 3 and imaging len 8, start laser instrument 4, this laser instrument 4 is the work light of detecting device, places between condenser lens 3 and the imaging len 8.
Inject the fluorescent reagent methylene blue, fine tuning micromatic setting 20 makes LASER SPECKLE be positioned at the center of sample cell 1 just again, adjusts diaphragm 11 the fluorescence spot is just passed through.
Adjustable diaphragm 11 adopts the mode of macropore 28 and aperture 29 stacks, assurance is in the diaphragm 11 picture that correctly focuses on detection position LASER SPECKLE just, can be detected by diaphragm 11, and fluorescence that sample cell 1 other position produces or parasitic light are owing to can not focus on and conductively-closed at diaphragm 11 places, can significantly reduce background stray light like this, obtain good signal to noise ratio (S/N ratio).
Effective diaphragm depends on aperture 29, and its structure is seen Fig. 4 a and Fig. 4 b.The present invention has been equipped with the diaphragm of the big small-bore of a series of differences, adopt different apertures at different systems, but for a certain system, pore size is fixed.
Fig. 2 b is that laser instrument 4 starts back sample cell 1 imaging in sight glass 9, pore wherein is a fluorescence spot imaging, the fixed knob 26 that screws after adjustment finishes on the micromatic setting 20 locks sample cell position 22, takes out sight glass 9, closes secondary light source 2 and weakener 5.Because sample cell 1 is to place replaceably on the micromatic setting 20, so after micromatic setting 20 calibrations and the locking, in continued operation, changes sample cell 1 and can not have influence on calibrated light path.
After finishing the light path set-up procedure, just can test sample, flow out component from the Micro-HPLC separating column and directly introduce sample cell 1 by polyfluortetraethylene pipe (Teflon), photomultiplier 12 output signals are connected to workstation or registering instrument record chromatographic peak signal, finish detection sample.Photodiode 7 plays the reference effect, and the output pulsation of laser instrument 4 is compensated.Because separating column will flow out the known technology that is connected to that component is introduced sample cell and photomultiplier and registering instrument by pipeline, the present invention is not described further at this, does not recommend accompanying drawing yet.
Compact conformation of the present invention only need change corresponding semiconductor laser 4 and optical filter 10 at different analysis systems, and these two parts can be changed at an easy rate all,
Embodiment given below can illustrate effect of the present invention:
The portable laser induced fluorescent tester volume of present embodiment is 13 * 30 * 9cm only, weighs about 5 kilograms;
The wavelength of semiconductor laser is 635nm, 3mw;
One section fused silica kapillary of detection cell: 0.25mmi.d, one section 3mm left and right sides of burning-off polyimide coating is as detection window;
Chromatographic condition: flow velocity: 8 μ l/min;
Moving phase: second cyanogen/acetate buffer solution (0.1M, PH=4.5), 60/40 (V/V);
Sample size: 100nl;
Separating column: the C of 0.32mm i.d * 9cm
18Post;
With methylene blue reagent as detection material.
Testing result: detect and be limited to 0.2ppb, quality testing is limited to 20amole, and signal to noise ratio (S/N ratio) (S/N) is 3: 1.Preparation 0.2ppb～0.2ppm series of standards solution, peak height and concentration are good linear relationship, and the range of linearity is 0.2ppb～20ppb, coefficient R=0.9976 (N=5).Figure five is the methylene blue chromatogram of 5ppb, has done a series of experiments under this concentration, repeatability fine (RSD＜3%, n=3).
Claims (2)
1, a kind of laser induced fluorescence detector includes:
One sample cell places the front end of a lens combination;
One lens combination is made up of condenser lens, imaging len;
One diaphragm and an optical filter place between lens combination and the photomultiplier, and wherein diaphragm places the lens combination rear side, and optical filter places between diaphragm and the photomultiplier;
One laser instrument is the work light of detecting device, places between condenser lens and the imaging len;
Induce the fluorescer of sample cell to produce fluorescence by above-mentioned laser instrument, be gathered in the photomultiplier output detection signal, it is characterized in that, also include through the lens combination imaging:
One is used to adjust the secondary light source of light path, is arranged between sample cell and the condenser lens;
One sight glass on the observation station between diaphragm and the optical filter, is used for the observation optical path adjustment process;
One weakener is arranged on the light path of laser instrument, between condenser lens and imaging len;
Described sample cell places on the micromatic setting, this micromatic setting has a slideway base, cross sliding clock and vertical slipper are installed on the base, the middle part is provided with the circular hole of a placement sample cell, and be provided with and move horizontally bar, move horizontally bar by adjustment cross sliding clock and vertical slipper are slided at slideway base, make sample cell do moving up and down within the specific limits, a fixed knob is used to lock the sample cell position;
The adjustable diaphragm that described diaphragm is made up of a macropore diaphragm and aperture stack.
2, laser induced fluorescence detector according to claim 1 is characterized in that, described sample cell is for being placed on the micromatic setting replaceably.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB011238623A CN1167945C (en) | 2001-08-07 | 2001-08-07 | Laser induced fluorescence detector |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB011238623A CN1167945C (en) | 2001-08-07 | 2001-08-07 | Laser induced fluorescence detector |
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| CN1400464A CN1400464A (en) | 2003-03-05 |
| CN1167945C true CN1167945C (en) | 2004-09-22 |
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| CNB011238623A Expired - Fee Related CN1167945C (en) | 2001-08-07 | 2001-08-07 | Laser induced fluorescence detector |
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Families Citing this family (13)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1316244C (en) * | 2004-02-26 | 2007-05-16 | 复旦大学 | Multi-chromaticity-electrophoresis separating and detecting system based on array capillary electrofocasing |
| JP2006134957A (en) * | 2004-11-02 | 2006-05-25 | Nidec Sankyo Corp | Manufacturing method of optical instrument |
| CN100590422C (en) * | 2005-11-06 | 2010-02-17 | 中国科学院安徽光学精密机械研究所 | Water body pollution laser induction fluorescence remote-measuring method |
| JP5272965B2 (en) * | 2009-08-19 | 2013-08-28 | 株式会社島津製作所 | Fluorescence detector |
| CN103514966B (en) * | 2012-06-20 | 2016-04-20 | 同济大学 | The sighting device of plasma diagnosis X-ray optical system and method for sight |
| CN103308503B (en) * | 2013-06-06 | 2015-11-25 | 四川大学 | Be separated microtrabeculae coupling light transmitting fiber exciting light induced fluorescence assembly |
| CN103529005B (en) * | 2013-10-16 | 2017-02-15 | 江苏禾尔思生物科技有限公司 | Application of high-efficiency capillary electrophoresis-laser induced fluorescence detection device and diagnostic kit in early diagnosis of mammary cancer |
| CN104749299B (en) * | 2013-12-25 | 2016-08-24 | 中国科学院大连化学物理研究所 | A kind of large volume flow cell for liquid chromatogram fluorescence detector |
| CN104483297B (en) * | 2014-12-08 | 2017-03-08 | 上海通微分析技术有限公司 | Visualization calibration laser-Induced Fluorescence Detection equipment |
| CN104849444B (en) * | 2015-05-20 | 2016-11-30 | 大连海事大学 | Fluorescence and block the cell counter and method simultaneously measured |
| CN106290208A (en) * | 2016-07-28 | 2017-01-04 | 青岛海纳光电环保有限公司 | A kind of ozone concentration determinator |
| CN110591904A (en) * | 2019-09-25 | 2019-12-20 | 福州大学 | Device for detecting miRNA |
| CN116879240B (en) * | 2023-09-07 | 2023-12-05 | 深圳市启扬光学科技有限公司 | Optical crystal detection system |
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