CN102297854B - High-efficiency multi-mode laser-induced fluorescence optical path exciting system - Google Patents
High-efficiency multi-mode laser-induced fluorescence optical path exciting system Download PDFInfo
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Abstract
The invention provides a high-efficiency multi-mode laser-induced fluorescence optical path exciting system. The system comprises a laser emitting apparatus, an excited apparatus, a light-splitting apparatus, a photoelectric converting apparatus, and a signal processing apparatus. The system is characterized in that: 1/2 wave plates are arranged on an optical path formed by a collimated beam which is emitted from the laser emitting apparatus, an optical path formed by a polarized-split beam, or split optical paths formed by emitted beams split by a plurality of polarized lights. With the exciting system, the polarization state of the excitation light is consistent with the polarization selectivity direction of the excited system. Within a same power, the signal intensity of excited Raman fluorescence is improved by 20% to 50%. When original polarization state compatibility is poor, with the system provided by the invention, the signal intensity can be improved by 50% to 80% or higher.
Description
Technical field
The present invention relates to a kind of fluorescence induction device, be specifically related to a kind of efficient multi-modal laser-induced fluorescence (LIF) light path activating system.
Background technology
Laser-induced fluorescence (LIF) light path activating system is mainly used in bioanalysis and detection ranges such as protein, DNA, and the vitochemical spectrometric instrument equipment of macromolecular mass.
As shown in Figure 1, the ultimate principle of its light path device and structure are: laser instrument 1 produces laser beam, after filtration after the mating plate 2, assemble through convergent lens 3, enter by by incidence window 4, the be excited container 5 of material of loading, material 6 is excited, the system that is excited that outgoing window 7 is formed, after the material 6 that is excited absorbs laser energy, energy level transition is sent fluorescence or Raman light 8, process is by collimation lens set 9, beam splitter 10, the beam splitting system that imaging lens group 11 grades are partly formed is come light splitting, the spectrum 12 of different wave length separates according to the locus, focal imaging is to photoelectric sensor 13, after photoelectricity transforms, the electric signal of the certain signal to noise ratio (S/N ratio) that obtains is transferred to back-end system and carries out analyzing and processing.
Wherein, laser instrument 1 refers to container 5 that multi-modal laser instrument loads the material that is excited the normally quartz, organic glass etc. of transparent material, and material requires the effective fluorescence signal that is excited out or the spectrum of Raman light signal are seen through preferably.Photoelectric sensor 13 is low light level signal transducers, and photomultiplier, CCD or photodiode are arranged usually.The material 6 that is excited has DNA, protein, big molecule organic chemicals or the material to be detected that can evenly float on a liquid etc. usually.
The laser-induced fluorescence (LIF) device that improves in the patent of the present invention has following 2 kinds of patterns usually: the burnt light channel structure pattern of copolymerization and orthogonal optical line structure pattern.The technical characterstic of copolymerization close-burning configuration formula is: excitation light path is identical with the optical axis direction that detects light path.Orthohormbic structure type technology characteristics are: the laser incident direction is vertical with the fluoroscopic examination direction or become certain angle between 0～90 °.
The light path of these two kinds of patterns, all be subjected to the light source polarizability and the influence of system's polarizability to launching efficiency that be excited: the laser that laser instrument sends, usually has certain polarization state (polarization direction), and the system that is excited (the especially carrying container of macromolecular organic compound matter, crystalline material making) also has certain polarization direction selectivity usually, if two kinds of polarizabilities are inconsistent, the absorption efficiency of material of being excited will be in various degree reduction, the fluorescence that ejects thus or Raman light signal intensity will not reach the highest stimulation effect.
The fluorescence that ejects or the intensity of Raman signal usually than a little less than the exciting light many, for example fluorescence intensity have only excitating light strength usually one of percentage to ten thousand/intensity, and Raman light have only usually ten thousand of excitating light strength/to 1,000,000/.Therefore, effectively promoting launching efficiency, promote signal to noise ratio (S/N ratio), is the Key Performance Indicator of laser-induced fluorescence detection system.
Usually the optimization method of attempting to eliminate this polarization selectivity has two kinds:
First kind is by the quarter wave plate of specific wavelength being set, linearly polarized light or elliptically polarized light process being adjusted to circularly polarized light, thereby reducing polarized light to the influence of the system of being excited.Its shortcoming is the polarization direction selectivity that can not influence the system of being excited 4, and the laser energy that 4 pairs of the systems that is excited meet its polarization choice direction has absorption preferably, and is still low to the absorption efficiency of the inconsistent portion of energy in polarization direction.
Second kind is the polarization direction of accurate measuring laser beam, by the method for regulating structure of precision, that the polarization choice direction of the system of being excited is consistent with the polarization direction of laser beam.The difficult point of this kind optimization method is, for multi-modal laser instrument product, and the polarization direction of its outgoing beam, and the multi-wavelength's of same laser device polarization direction, always not consistent; Simultaneously, for need carrying out in the light path design of beam splitting laser instrument, the polarization spectro sheet that manufactures and designs under the current techniques condition always can only accurately carry out beam splitting to S light component and the P light component of the some specific wavelengths in the middle of the multi-wavelength of multi-modal laser emitting.This difficulty especially is embodied in the batch process of product, and the designer lacks operability to the situation that each situation designs and produces separately to reach optimum.
Summary of the invention
At above-mentioned defective, the purpose of this invention is to provide a kind of efficient multi-modal laser-induced fluorescence (LIF) light path activating system, to solve the low technical matters of laser-induced fluorescence (LIF) light path device launching efficiency of prior art.
For achieving the above object, the present invention has adopted following technical scheme:
A kind of efficient multi-modal laser-induced fluorescence (LIF) light path activating system, comprise laser beam emitting device, the device that is excited, light-dividing device, photoelectric conversion device and signal processing apparatus, described laser beam emitting device send light beam on the light path behind the collimation, described emission light beam through on the light path behind the polarization spectro or the light splitting light path of described emission light beam after a plurality of polarized lights are divided into multi-beam be provided with 1/2 wave plate.
According to the described activating system of preferred embodiment of the present invention, described laser beam emitting device is multi-modal laser instrument, its emission light is after an optical filter filters, change optical path direction again through a polarization spectro sheet light splitting through first catoptron, part light after the light splitting is injected described device one side that is excited after through second mirror reflects, another part light is through the 3rd of two correspondences the, inject from the described device opposite side that is excited behind the 4th catoptron, two bursts of light paths excite the material that is excited in the described device that is excited to send fluorescence or Raman light, focus on described photoelectric conversion device through light-dividing device, obtain electric signal and be transferred to described signal processing apparatus, between described polarization spectro sheet and described second catoptron, and the described the 3rd, all be provided with 1/2 wave plate between the 4th catoptron.
According to the described activating system of preferred embodiment of the present invention, described 1/2 wave plate is used for the polarization mode of the excitation beam specific wavelength of the described laser beam emitting device of adjustment, makes that the polarization direction of light beam is consistent with the polarization selectivity direction of the described device that is excited.
According to the described activating system of preferred embodiment of the present invention, the described device that is excited comprises that further one is provided with the container of transparent window, and this container contents is equipped with the material that is excited.
According to the described activating system of preferred embodiment of the present invention, described the 4th catoptron and described being excited between the device, and described second catoptron and described being excited respectively are provided with one and assemble lens between the device.
According to the described activating system of preferred embodiment of the present invention, described light-dividing device comprises collimation lens set, grating and imaging lens group, the light direction that described collimation lens set is arranged on the described device that is excited through the grating adjustment after light inject and be imaged onto described photoelectric conversion device after described imaging lens group focuses on.
According to the described activating system of preferred embodiment of the present invention, described 1/2 wave plate is installed on the wave plate installation adjusting device, described wave plate installation adjusting device comprises mount pad, be installed in the erecting frame that described mount pad is provided with runner, the tight retainer of wave plate frame and the wave plate pressure ring that is used for locking described erecting frame, and described 1/2 wave plate is arranged on the described erecting frame by described wave plate pressure ring and several packing rings.
Owing to adopted above technical characterictic, make the present invention than prior art, have following advantage and good effect:
The present invention is by in the light path of multi-modal laser, any position on the light path under to the situation of single beam excitation after the collimation, to any position on the light path of laser beam after through light splitting under the situation after the light splitting of polarization spectro sheet or laser beam is divided into any position on the light path after the light splitting under the situation of multi-beam through a plurality of polarization spectro sheets, some 1/2 wave plates are set adjust the polarization mode of excitation beam specific wavelength (seeing accompanying drawing five), thereby making excites the polarization selectivity direction of polarization state of light and the system of being excited consistent, with making the Raman fluorescence signal intensity that excites improve 20%～50% under the constant power, when original polarization state matching is relatively poor, can promote 50%～80% even higher.
Description of drawings
Fig. 1 is laser-induced fluorescence (LIF) device ultimate principle structural drawing;
Fig. 2 is 1/2 wave plate polarization theory figure;
Fig. 3 is wave plate installation adjusting device exploded view provided by the invention;
Fig. 4 is the Organization Chart of an embodiment of the present invention.
Embodiment
Below in conjunction with accompanying drawing several preferred embodiments of the present invention is described in detail, but the present invention is not restricted to these embodiment.The present invention is contained any in substituting of making of marrow of the present invention and scope, modification, equivalent method and scheme.Understand for the public is had completely the present invention, in the following preferred embodiment of the present invention, describe concrete details in detail, and do not have the description of these details also can understand the present invention fully for a person skilled in the art.
Core concept of the present invention is: by in the light path of multi-modal laser, in place, suitable wave plate device (a slice or multi-disc) is set adjusts the polarization mode of excitation beam specific wavelength, thereby making excites the polarization selectivity direction of polarization state of light and the system of being excited consistent, with making the Raman fluorescence signal intensity that excites improve 20%～50% under the constant power, when original polarization state matching is relatively poor, can promote 50%～80% even higher.
To the situation of single beam excitation, be arranged on any position on the collimation light path afterwards; Through the situation after the light splitting of polarization spectro sheet, the polarization spectro sheet is arranged on any position on the light path after the light splitting to laser beam; Laser beam is divided into the situation of multi-beam through a plurality of polarization spectro sheets, and the polarization spectro sheet is arranged on any position on the light path after the light splitting.
As shown in Figure 2, the ultimate principle of 1/2 wave plate, 37 deflection direction of vibration is ultimate principles of optics specialty, does not do tired stating herein.Key concept is summarized as follows: incident light is monochromatic light, and its polarized component is that S component (direction of vibration such as figure four) and P component (direction of vibration such as figure four) determine, when having only the S component or having only the P component, is called linearly polarized light.The light beam that includes S component and P component is elliptically polarized light.Effect to a wave plate in two fens is that change polarization direction 90 degree namely as the direction of vibration of figure emergent light and the direction of vibration of incident light, differ 90 degree.
As shown in Figure 3,1/2 wave plate among the present invention fixes the position by following wave plate installation adjusting device usually.The mentioned wave plate installation adjusting device of the present invention comprise wave plate pressure ring 38, rubber washer 39,21,2 wave plates 40, packing ring 41, wave plate erecting frame 42, wave plate mount pad 43, the tight retainer 44 of wave plate frame, tightly lock screw 46, tightly lock screw 50 etc. and partly form.
Wherein, the effect of wave plate pressure ring 38 is to be fastened on uniformly on the wave plate erecting frame wave plate is stressed, and the annexation fine thread between wave plate pressure ring and the wave plate erecting frame is fastening, can coat screw thread glue during permanent installation.Wave plate erecting frame 42 is installed in the wave plate mount pad circular hole as shown in the figure, tolerance on fit+0.01～+ 0.05mm, can rotate freely, but the rolling momentum is again little.The setting height(from bottom) of wave plate mount pad 43 is determined by the height of real system optical axis distance installed surface.The effect of the runner 48 of wave plate erecting frame 42 is manually or by the accurate rotating wave plate angle of other frocks, thereby obtains the optimum efficiency that will reach mentioned above.Tight effect of locking screw 46 is after debugging is finished, to make the tight retainer 44 of wave plate erecting frame and wave plate erecting frame 42 tighten together by screw 49, the wave plate position that minimizing causes owing to vibrations, angle variation.The effect of tight lock screw 50 is to make wave plate erecting frame 43 and wave plate erecting frame 42 tightly lock, and is installed in screwhole position 45.
Usually, light path is generally about 1～3 ° the angle sensitivity of wave plate, and this structure adopts the wave plate erecting frame 42 (runner diameter is greater than 2 times of wave plate diameters) of band big roller.Wave plate diameter 12mm for example, roller diameter is more than 24mm so, and adjustment amount is at 0.5mm～1.5mm.This runner of free-hand rotation relies on the perception of hand, namely can reach this precision.Also can design the little rod 47 of such adjusting, on roller, (see 48 holes of arranging, the diameter 2mm in hole every circular hole of 30 degree processing, degree of depth 5mm, totally 12), during adjusting, one section of little rod frock 47 is inserted in the hole, holds an other end, regulates the wave plate angle by handle.Little rod frock 47 can design about 50mm～100mm, and the angle orientation scope that adjust this moment can reach 1.7mm～5.1mm, and the angle orientation ratio of precision said method that can reach is high more than 2～4 times.
Please refer to Fig. 4, discuss the synoptic diagram of the present invention's one specific embodiment, it comprises laser instrument 51, after filtration after the mating plate 52, change optical path direction via catoptron 53, through polarization spectro sheet 54, via catoptron 55,56,57 change optical path direction, through the focal length convergent lens 58 identical with relative aperture, 59, enter by loading the be excited container 60 of material and material 61 devices that are excited dimerous that are excited, be excited and send fluorescence or Raman light 62 behind the material 61 absorption portion laser energies, design transparent window 63 usually on loading is excited the container 60 of material, through comprising collimation lens set 64, the light-dividing device of beam splitter such as grating 65 imaging lens group 66, focal imaging are to photoelectric sensor 67, thereby the electric signal that obtains certain signal to noise ratio (S/N ratio) is transferred to and carries out analyzing and processing in the signal processing apparatus 66.
The laser instrument of mentioning in this example 51 refers to multi-modal laser instrument.Loading be excited the container 60 normally quartz, organic glass etc. of transparent material of material 61, material requires the effective fluorescence signal that is excited out or the spectrum of Raman light signal are seen through preferably.Photoelectric sensor 67 is low light level signal transducers, and photomultiplier, CCD or photodiode are arranged usually.The material 61 that is excited has DNA, protein, big molecule organic chemicals or the material to be detected that can evenly float on a liquid etc. usually.In this example, adopting the purpose of incident double light path in opposite directions, is to make the stimulation effect uniformity at two ends.Because container 60 is bigger in the length of optical axis 69 directions, the luminous energy on monochromatic light road is when penetrating material, and energy reduces gradually, can significantly descend to other one section stimulation effect, therefore, adopts in opposite directions that double light path excites simultaneously, to obtain sufficient stimulation effect.Simultaneously, known detected system has polarization selectivity (container or the detected material that namely are contained in detected material matter have certain polarization selectivity).Detected material has 2 kinds of Raman light labeling dyes, is respectively the 6-FAM (maximum absorption wavelength 494nm, maximum emission wavelength 518nm) of Japanese TaKaRa company, HEX (maximum absorption wavelength 533nm, maximum emission wavelength 559nm).
Laser instrument is argon ion gas laser.Laser power 50mW, beam diameter 1mm includes 458nm, 476nm, 488nm, 497nm, 502nm, 514.5nm Deng 6 main spectral lines, wherein the energy of 488nm and 514.5nm spectral line occupies more than 80% of gross energy, wherein 488nm energy and 514.5nm energy ratio are about 2: 1.
The method that patent of the present invention is related is placed 2 identical 1/2 wave plates 70,71 by adjustment erecting device shown in Figure 3 in light path device.
The reason of placing this position has: (1) is in full accord with respect to light path component number and the optical performance parameter at the material two ends that are excited, and it is just the same that wave plate is placed this location status, and the position has the space interchangeability; (2) light path of this position is longer, is suitable for arranging on the structure wave plate structural member, can leaving space to the debugging of wave plate.
Laser power is 50mW, and through after the light splitting, the maximum energy-density on each wave plate surface is 50000w/m
2, should adopt the high Coating Materials titania (TiO of tolerance emittance
2) (this material tolerance energy density is up to 2x10 as the anti-reflection film material
6W/m
2More than, these data are for disclosing inspection information), and be suitable for long-term work.
1/2 wave plate that the related method of patent of the present invention is carried if system pays the utmost attention to the Raman light that the 6-FAM dyestuff sends, then can design 1/2 wave plate of 488nm wavelength location; If system pays the utmost attention to the Raman light that the HEX dyestuff sends, then can design 1/2 wave plate of 514.5nm wavelength location.If balance is taken into account two kinds of dyestuffs, the wavelength of 1/2 wave plate should be between 488nm and 514.5nm.
Can calculate its efficient as table 1.
Table 1
X11 in the last table, X12, X21, the X22 data are measured data of experiment.Only for illustrating the use of method, only for reference.In the formula (2) that the substitution summary of the invention is the 2nd, (3), (4), the numerical value of the characteristic wavelength λ that can calculate is 501.25nm, i.e. the wavelength design load of 1/2 wave plate.
The preferred embodiment of the present invention just is used for helping to set forth the present invention.Preferred embodiment does not have all details of detailed descriptionthe, does not limit this invention yet and only is described embodiment.Obviously, according to the content of this instructions, can make many modifications and variations.These embodiment are chosen and specifically described to this instructions, is in order to explain principle of the present invention and practical application better, thereby the technical field technician can utilize the present invention well under making.The present invention only is subjected to the restriction of claims and four corner and equivalent.
Claims (6)
1. detection system that adopts efficient multi-modal laser instrument induced fluorescence excitation light path, comprise laser beam emitting device, device is excited, light-dividing device, photoelectric conversion device and signal processing apparatus, it is characterized in that, described laser beam emitting device is multi-modal laser instrument, its emission light is after an optical filter filters, change optical path direction again through a polarization spectro sheet light splitting through first catoptron, part light after the light splitting is injected described device one side that is excited after through second mirror reflects, another part light is through the 3rd of two correspondences the, inject from the described device opposite side that is excited behind the 4th catoptron, send fluorescence or Raman light from the material that is excited in the described device that is excited of the optical excitation of two bursts of light paths, collimate through light-dividing device, light splitting and focus on described photoelectric conversion device, obtain electric signal and be transferred to described signal processing apparatus, between described polarization spectro sheet and described second catoptron, and the described the 3rd, all be provided with 1/2 wave plate between the 4th catoptron.
2. detection system as claimed in claim 1 is characterized in that, described 1/2 wave plate is used for the polarization mode of the excitation beam specific wavelength of the described laser beam emitting device of adjustment, makes that the polarization direction of light beam is consistent with the polarization selectivity direction of the described device that is excited.
3. detection system as claimed in claim 1 is characterized in that, the described device that is excited comprises that further one is provided with the container of transparent window, and this container contents is equipped with the material that is excited.
4. detection system as claimed in claim 1 is characterized in that, described the 4th catoptron and described being excited between the device, and described second catoptron and described being excited respectively are provided with one and assemble lens between the device.
5. detection system as claimed in claim 1, it is characterized in that, described light-dividing device comprises collimation lens set, grating and imaging lens group, the light direction that described collimation lens set is arranged on the described device that is excited through the grating adjustment after light inject and be imaged onto described photoelectric conversion device after described imaging lens group focuses on.
6. detection system as claimed in claim 1, it is characterized in that, described 1/2 wave plate is installed on the wave plate installation adjusting device, described wave plate installation adjusting device comprises mount pad, be installed in the erecting frame that described mount pad is provided with runner, the tight retainer of wave plate frame and the wave plate pressure ring that is used for locking described erecting frame, and described 1/2 wave plate is arranged on the described erecting frame by described wave plate pressure ring and several packing rings.
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| CN103063640B (en) * | 2012-12-28 | 2015-09-09 | 西北核技术研究所 | A kind of laser-induced fluorescence (LIF) combustion field parameter measuring apparatus |
| CN106383082B (en) * | 2015-10-14 | 2018-10-19 | 北京信息科技大学 | A kind of optical path adjustment device and method of the flow cytometer without fluid path situation |
| CN105388140B (en) * | 2015-12-01 | 2017-05-10 | 杭州中车车辆有限公司 | Measuring instrument for site invisible fingerprint display and contained substance thereof |
| CN106290299A (en) * | 2016-08-04 | 2017-01-04 | 北京华泰诺安探测技术有限公司 | A kind of polarization diversity polarization Raman probe and optical spectrum detecting method |
| CN107449762B (en) * | 2017-08-15 | 2020-03-13 | 重庆三峡学院 | Tape-type monitoring method and equipment for early warning of water surface oil spill pollution |
| CN111060484B (en) * | 2019-12-29 | 2020-11-03 | 中国科学院西安光学精密机械研究所 | Non-scanning three-dimensional plane laser-induced fluorescence imaging detection method and system |
| CN116602628B (en) * | 2023-07-17 | 2023-11-28 | 江苏京泰全医疗科技有限公司 | Detection device for self-fluorescence tissue |
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| WO2002079762A2 (en) * | 2000-10-27 | 2002-10-10 | Dumas David P | Apparatus for fluorescence detection on arrays |
| JP2005345561A (en) * | 2004-05-31 | 2005-12-15 | Olympus Corp | Scanning type laser microscope device |
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