CN102305778B - Micro-multispectral fluorescence reception and treatment system - Google Patents
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- CN102305778B CN102305778B CN 201110128960 CN201110128960A CN102305778B CN 102305778 B CN102305778 B CN 102305778B CN 201110128960 CN201110128960 CN 201110128960 CN 201110128960 A CN201110128960 A CN 201110128960A CN 102305778 B CN102305778 B CN 102305778B
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Abstract
The invention discloses a micro-multispectral fluorescence reception and treatment system. The micro-multispectral fluorescence reception and treatment system comprises an excitation unit, a fluorescence collection unit and a data processing unit, wherein the excitation unit comprises an excitation light source, an exciting light filter and an optical convergence component; the fluorescence collection unit comprises a photoelectric converter and a micro-optical filter which is attached to the photoelectric converter closely; and the micro-optical filter is a highly integrated multi-channel optical filter. An object to be imaged radiates the fluorescence with a specific wavelength under the excitation of irradiation light of the excitation unit; the optical convergence component converges the fluorescence; then the converged fluorescence is irradiated to the photoelectric converter through the micro-optical filter; the photoelectric converter acquires corresponding image information according to a fluorescence signal; and the data processing unit analyzes and processes the image information and outputs a result. The system can collect multiple fluorescence signals of stimulated radiation and acquire image information in a high-efficiency and low-cost way, and can accurately and objectively reflect detection results in real time.
Description
Technical field
The present invention relates to biochemistry detection and external medical diagnostic equipment, relate in particular to a kind of little multispectral fluorescence reception and disposal system.
Background technology
At present, the employed instrument of medical institutions' clinical diagnosis and biological detection, (Picymerase Chain Reaction) PCR is feature all take the PCR in the fluorescent collecting system of employing.In the PCR reaction system, add the primer that contains fluorescent material, fluorescent material sends the light of another feature wavelength under the excitation source irradiation of specific wavelength, detect the light intensity that fluorescent material is launched in the PCR course of reaction by photomultiplier acceptance, optical fiber optical signal transmission, discrete optical filter light splitting, monitoring pcr amplification reaction process.Draw real-time analytic curve by computer software analysis, calculate the Ct value, provide the nucleic acid concentration value of sample starting template, carry out detection and the genetic analysis of the various pathogen of DNA/RNA.
It is discrete optical filter light splitting that existing clinical diagnosis, biochemical instrument adopt the method for designing of fluorescence gathering system, the optical fiber optical signal transmission, photomultiplier is accepted fluorescence signal and is carried out opto-electronic conversion, process post analysis calculating original gene content and expression formula through preposition amplification and data, detect the situation of sample tissue element variation.This project organization cost is high, and the collection of light and opto-electronic conversion are complicated, the poor stability of authentic specimen acquisition of information, and measurable number of samples is restricted.
Adopt common photomultiplier, to convert faint optical signal to electric signal, but when illumination is mapped to photomultiplier, photocathode inspires photoelectron in vacuum, photoelectron enters dynode system by the focusing board electric field, obtain multiplication by further Secondary Emission and amplify, the electronics after the amplification is collected with anode and is exported as signal.Owing to adopted the Secondary Emission dynode system, so photomultiplier has higher sensitivity in the photodetection of the emittance of ultraviolet, visible light and near-infrared region, be widely used in the clinical diagnosis of carrying out optical measurement and spectral analysis and biochemical instrument.But common photomultiplier once can only be measured an information, and passage only has one, has limited the measurement of multi-channel information; Photomultiplier in actual use, detection sensitivity reduces because of strong illumination or because irradiation time is long, stops after the irradiation again part and recovers, and " tired " phenomenon occurs; The sensitivity of photomultiplier photoanode surface each point is inhomogeneous.Photomultiplier also needs accessory circuit such as high-voltage power supply, amplification and Discr., cool room, magnetic shielding, base etc. in addition, and this is high to the requirement of environment just so that the volume of whole system is large.
In recent years, along with the development of semiconductor technology, multi-anode photomultiplier and silicon photomultiplier are come out one after another, and its common characteristics are to make many very thin photomultipliers form matrix, increase detection channels, reduce the volume of photomultiplier; Optical detection unit adopts the optical fiber lens arra.But the anode end of multi-anode photomultiplier is subjected to the restriction of fine wire, and it is measured passage and can only accomplish hundred at present.The optical fiber that is used for optical detection unit, for sample carriers, owing to be point measurement, the light signal of detection and transmission is the real information of complete representative tested sample accurately.Aspect the fixed form of optical fiber: optical fiber is fixed on the bottom of sample, if the location will definitely not affect the detection quality of signal; Optical fiber is fixed on the top of sample, and in the experimentation, the movement that microwell plate can be frequent not only affects the quality of measurement but also brings inconvenience to the operator, simultaneously optical fiber contaminated accuracy that also can reduce measurement in experimentation.
Adopt discrete optical filter light splitting, in a large amount of experimentations, need frequent adjustment optical filter wheel to make the discrete different wavelength of optical filter light splitting, so not only can produce artificial noise, because the Mechanical Moving of optical filter wheel can cause the position skew of obtaining image, the quality of image information is obtained in impact simultaneously.
Summary of the invention
The technical matters that the present invention mainly solves provides a kind of little multispectral fluorescence reception and disposal system, overcoming existing fluorescence reception and processing unit can not be in real time, accurately, the defective of the real information that obtains fluorescence signal of low-cost, miniature multi-channel.
For solving the problems of the technologies described above, the technical scheme that the present invention adopts is: a kind of little multispectral fluorescence reception and disposal system are provided, comprise: for generation of monochromatic light uniform irradiation object to be imaged with excite object radiation fluorescence to be imaged excite the unit, be used for collecting the fluorescence of object radiation to be imaged and be converted into the phosphor collection unit of image information and to carry out the data processing unit of analyzing and processing according to the image information of fluorescence conversion; Described phosphor collection unit comprises that optics converges parts, photoelectric commutator and is close to the micro optical filter of photoelectric commutator input end, described micro optical filter is highly integrated multi-channel filter, described micro optical filter comprises a plurality of compact arranged optical filtering infinitesimals, described optical filtering infinitesimal is matrix to be arranged, and a described optical filtering infinitesimal covers a photoelectric conversion unit of described photoelectric commutator; Described optics converges parts and is arranged on the light path of object fluorescent radiation to be imaged, and described micro optical filter is arranged on the reflected light path that described optics converges parts, and the output terminal of described photoelectric commutator connects described data processing unit; Wherein, it is concave mirror that described optics converges parts, and described micro optical filter reaches integrated with it photoelectric commutator and is arranged on the focus of described concave mirror; Describedly excite irradiation that the unit launches on object to be imaged, object to be imaged is stimulated and gives off the fluorescence of specific wavelength, fluorescence is radiated at described optics and converges on the parts, optics converge that parts converge to micro optical filter with fluorescence and the photoelectric commutator that fits tightly with micro optical filter on, micro optical filter allows the fluorescence signal of specific wavelength to pass through, fluorescence signal is radiated on the photoelectric commutator after by micro optical filter, photoelectric commutator obtains corresponding image information according to described fluorescence signal, and sending image information to described data processing unit, data processing unit is demarcated the information that described photoelectric commutator sends, analyze and process and export the result of analyzing and processing; Described data processing unit is Computerized image processing system, the image information that described photoelectric commutator obtains is delivered to demarcation and the processing that Computerized image processing system carries out image by Computer interface circuit, treat imaging object and carry out real-time analysis and quantitatively detect, and the result of analyzing and processing is transported on the graphoscope shows.
Wherein, the photoelectric commutator of described phosphor collection unit is CCD or cmos image sensor, and described micro optical filter is integrated on CCD or the cmos image sensor.
Wherein, the photoelectric commutator of described phosphor collection unit is for to press the compact arranged photomultiplier of array, and micro optical filter is integrated on the photomultiplier of little gust of arrangement.
Wherein, the described unit that excites comprises LASER Light Source and convex reflecting mirror, and described convex reflecting mirror is arranged on the emitting light path of LASER Light Source, and described object to be imaged is arranged on the reflected light path of convex reflecting mirror.
Wherein, the described unit that excites comprises arrowband high-power LED light source and exciting light optical filter, is radiated at equably on the object to be imaged behind the emergent light process exciting light optical filter of described arrowband high-power LED light source.
Wherein, the described unit that excites comprises arrowband high-power LED light source, exciting light optical filter and plane mirror, the emergent light of described arrowband high-power LED light source shines described plane mirror through the exciting light optical filter, and described object to be imaged is positioned on the reflected light path of described plane mirror.
The invention has the beneficial effects as follows: be different from the clinical diagnosis of prior art, fluorescence reception in the biochemical instrument and disposal system adopt photomultiplier to accept, the optical fiber optical signal transmission, discrete optical filter light splitting is so that image is inaccurate, huge structure, high and the testable limited amount of cost, little multispectral fluorescence reception of the present invention and disposal system adopt miniature multispectral technology, utilize micro optical filter that the fluorescence signal of the radiation that is stimulated of collection is filtered, micro optical filter is highly integrated multi-channel filter, be close to the input end of photoelectric commutator, arrowband fluorescence signal corresponding to the primary fluorescence specific wavelength obtains corresponding image by being radiated at photoelectric commutator from the microchannel of micro optical filter, Output rusults after processing through data processing unit.In the present invention, for the fluorescence signal of reporting the biochemical reaction process that the fluorescent dye sample occurs, become on the whole real-time little multispectral image transmission, and then recorded by the photoelectric commutator with micro optical filter, the image that obtains accurately, sensitive, reflect the Biochemical Information of each test tube sample carrier in the micropore template objectively, improved the signal to noise ratio (S/N ratio) of image and detected the reliability of sample.Only needing to adjust miniature narrow band pass filter allows just to go for the fluorescence reception of different series and clinical diagnosis and the biochemical instrument of processing procedure by light wavelength.Biological detection and clinical diagnosis instruments such as fluorescent colloid detector, fluorescent polyase chain reaction instrument, immune time resolution detector, immunochemiluminescence detector.The present invention makes instrument easily, the mode of collecting fluorescence designs very compact, succinct, be easy to simultaneously standardization, the reliability of product reaches high standard easily, quality in the instrument manufacturing process is controlled easily, the manufacturing at product has reduced cost, the complexity decrease of design difficulty and instrument.
Description of drawings
Fig. 1 is the light path principle figure of one embodiment of the invention;
Fig. 2 is the structured flowchart of embodiment among Fig. 1 of the present invention;
Fig. 3 is the structural representation of micro-filtration wave plate of the present invention;
Fig. 4 is A place partial enlarged drawing among Fig. 3;
Fig. 5 is the light path principle figure that the present invention uses LASER Light Source;
Fig. 6 is the index path that the present invention uses the arrowband high-power LED light source;
Fig. 7 is the index path that the present invention uses another embodiment of arrowband high-power LED light source.
Embodiment
By describing technology contents of the present invention, structural attitude in detail, realized purpose and effect, below in conjunction with embodiment and cooperate that accompanying drawing is detailed to give explanation.
See also Fig. 1 and Fig. 2, the invention provides a kind of little multispectral fluorescence reception and disposal system, comprising: excite unit 1, photoelectric conversion unit 2 and data processing unit 3, containing and being loaded with fluorescent dye 4 is that the test tube of object to be imaged is placed on the micropore template 5; Excite unit 1 to launch the monochromatic light uniform irradiation on the fluorescent dye 4 of micropore module 5, fluorescent dye 4 is stimulated and gives off the fluorescence of specific wavelength.Described phosphor collection unit 2 comprises that optics converges parts 23, photoelectric commutator 22 and is close to the micro optical filter 21 of photoelectric commutator 22 input ends, described micro optical filter 21 is highly integrated multi-channel filter, optics converges parts 23 and is arranged on the light path of fluorescent dye 4 fluorescent radiations, described micro optical filter 21 is arranged on the reflected light path that described optics converges parts 23, and the output terminal of described photoelectric commutator 22 connects described data processing unit 3.
Among the present invention, excite irradiation that unit 1 launches on fluorescent dye 4, fluorescent dye 4 is stimulated and gives off the fluorescence of intrinsic specific wavelength, fluorescence is radiated at described optics and converges on the parts 23, optics converges parts 23 fluorescence is converged on the micro optical filter 21, fluorescence signal is radiated at after by micro optical filter 21 on the photoelectric commutators 22 of closely placing with micro optical filter 21, in the present embodiment, optics converges parts 23 and adopts concave mirror, micro optical filter 21 and be arranged on the focus of concave mirror with photoelectric commutators 22 that micro optical filter 21 is closely placed, the fluorescence of fluorescent dye 4 radiation is radiated on the concave mirror, concave mirror converges fluorescence according to optical principle and increases light intensity, the fluorescence that reflects impinges upon on the micro optical filter 21, described micro optical filter 21 is highly integrated multi-channel filter, only allow the fluorescence signal of specific wavelength to pass through, photoelectric commutator 22 obtains corresponding image information according to fluorescence signal, and flowing to data processing unit 3, the information that 3 pairs of described photoelectric commutators 22 of data processing unit send is demarcated, analyze and process and export the result of analyzing and processing.Described data processing unit 3 can be Computerized image processing system, the image information that described photoelectric commutator 22 obtains is delivered to demarcation and the processing that Computerized image processing system carries out image by Computer interface circuit, fluorescent dye 4 is carried out real-time analysis and quantitatively detects, and the result of analyzing and processing is transported on the graphoscope shows.
Be different from the clinical diagnosis of prior art, fluorescence reception in the biochemical instrument and disposal system adopt photomultiplier to accept, the optical fiber optical signal transmission, discrete optical filter light splitting is so that image is inaccurate, huge structure, high and the testable limited amount of cost, little multispectral fluorescence reception of the present invention and disposal system adopt miniature multispectral technology, utilize micro optical filter that the fluorescence signal of the radiation that is stimulated of collection is filtered, micro optical filter is highly integrated multi-channel filter, be close to the input end of photoelectric commutator, arrowband fluorescent signal corresponding to the primary fluorescence specific wavelength obtains corresponding image by being radiated at photoelectric commutator from the microchannel of micro optical filter, Output rusults after processing through data processing unit.In the present invention, for the fluorescence signal of reporting the biochemical reaction process that the fluorescent dye sample occurs, become on the whole real-time little multispectral image transmission, and then recorded by the photoelectric commutator with micro optical filter, the image that obtains accurately, sensitive, reflect the Biochemical Information of each test tube sample carrier in the micropore template objectively, improved the signal to noise ratio (S/N ratio) of image and detected the reliability of sample.Only needing to adjust miniature narrow band pass filter allows just to go for the fluorescence reception of different series and clinical diagnosis and the biochemical instrument of processing procedure by light wavelength.Biological detection and clinical diagnosis instruments such as fluorescent colloid detector, fluorescent polyase chain reaction instrument, immune time resolution detector, immunochemiluminescence detector.The present invention makes instrument easily, the mode of collecting fluorescence designs very compact, succinct, be easy to simultaneously standardization, the reliability of product reaches high standard easily, quality in the instrument manufacturing process is controlled easily, the manufacturing at product has reduced cost, the complexity decrease of design difficulty and instrument.
Consult Fig. 3, micro optical filter 21 of the present invention comprises a plurality of compact arranged optical filtering infinitesimals 211, described optical filtering infinitesimal 211 is matrix and arranges, the size of described optical filtering infinitesimal must not be significantly less than the single Pixel Dimensions of corresponding described photoelectric commutator 22, and a described optical filtering infinitesimal 211 covers a photoelectric conversion unit of described photoelectric commutator 22.The arrowband fluorescent signal of primary fluorescence specific wavelength is by optical filtering infinitesimal 211 imaging on photoelectric commutator 22, between pixel and the pixel, automatically calibrating aligns between image and the image, make the image that obtains accurately, sensitive, reflect the Biochemical Information of each test tube sample carrier in the micropore template objectively, improved the signal to noise ratio (S/N ratio) of image and detected the reliability of model.The light wavelength that described optical filtering infinitesimal allows to pass through can be set to identical or different, and for example in Fig. 4, it is that the light of λ 1, λ 2, λ 3 and λ 4 passes through that four optical filtering infinitesimals can allow respectively wavelength, detects when can realize like this different wave length fluorescence.
In one embodiment, the photoelectric commutator 22 of described phosphor collection unit 2 is CCD or cmos image sensor, described micro optical filter 21 is integrated on CCD or the cmos image sensor, be specially micro optical filter 21 directly is produced on the opto-electronic conversion sensitive element of a CCD/COMS monochrome image sensor with low light level imaging capability, corresponding to the fluorescent signal of primary fluorescence specific wavelength by behind the micro optical filter 21, imaging on the imaging surface of the CCD/COMS monochrome image sensor with low light level imaging capability.Described photoelectric commutator 22 adopts CCD or COMS imageing sensor, and the CCD or the cmos image sensor that are integrated with micro optical filter 21 just become multispectral CCD/COMS camera or video camera.
In another embodiment, described photoelectric commutator 22 is for to press the compact arranged photomultiplier of array, and described micro optical filter 21 is integrated in this by on the compact arranged photomultiplier of array.Described micro optical filter 21 directly is produced on the photosensor chip in micro production mode (such as vacuum coating and microlithography technology etc.), and is integrated in the photomultiplier transit tube module.
Consult Fig. 5, in one embodiment, laser cell 1 comprises LASER Light Source 11 and convex reflecting mirror 14, convex reflecting mirror 14 is arranged on the emitting light path of LASER Light Source 11, the irradiation that LASER Light Source 11 is sent is on convex reflecting mirror 14, and light is radiated on the fluorescent dye 4 after convex reflecting mirror 14 reflections equably.In this embodiment, the light that LASER Light Source 11 is sent expands by convex reflecting mirror 14, convex reflecting mirror 14 has the characteristic to the incident light scattering, enlarged so catoptrical range of exposures, uniformly all standing shone each fluorescent dye 4 on the micropore template 5 when reflected light shone on the micropore template 5.
Consult Fig. 6, in another embodiment, excite unit 1 to comprise arrowband high-power LED light source 11a and exciting light optical filter 12, be radiated at equably on the fluorescent dye 4 behind the emergent light process exciting light optical filter 12 of described arrowband high-power LED light source 11a.Fluorescent dye 4 gives off fluorescence and converges parts 23 through optics and converge reinforcement, again by micro optical filter 21 imaging on photoelectric commutator 22.Imaging system 40 comprises that fluorescent dye 4 and optics converge parts 23 among Fig. 6, and the structure that it is concrete and light path be existing the specific descriptions above, herein to its simplification, no longer specifically describes.The imaging system that hereinafter occurs represents identical implication.
Fig. 7 has provided another embodiment as excitation source with arrowband high-power LED light source 11a, in this embodiment, excite unit 1 to comprise arrowband high-power LED light source 11a, exciting light optical filter 12 and plane mirror 13, the emergent light of arrowband high-power LED light source 11a incident angle with 45 ° after exciting light optical filter 12 filters is radiated on the plane mirror 13, the reflected light of plane mirror 13 shines on the imaging system 40, and the emergent light of imaging system 40 is radiated at imaging on the photoelectric commutator 22 by micro optical filter 21 again.When reality is used, reasonably adjust the angle of plane mirror 13 according to the position relationship of arrowband high-power LED light source 11a, micropore module 5, make irradiation that arrowband high-power LED light source 11a sends on plane mirror 13, fluorescent dye 4 is positioned on the reflected light path of plane mirror 13, and the light that arrowband high-power LED light source 11a sends can get final product through the reflection uniform fold fluorescent dye of plane mirror 13.
Among the present invention, produce the light source of exciting light except can for the LASER Light Source and arrowband high-power LED light source such as xenon lamp and so on, also using high-pressure sodium lamp, quartz-halogen-tungsten lamp or the LED laser instrument of co-wavelength scope as an alternative.Its embodiment repeats no more.
The above only is embodiments of the invention; be not so limit claim of the present invention; every equivalent structure or equivalent flow process conversion that utilizes instructions of the present invention and accompanying drawing content to do; or directly or indirectly be used in other relevant technical fields, all in like manner be included in the scope of patent protection of the present invention.
Claims (6)
1. a little multispectral fluorescence reception and disposal system is characterized in that, comprising:
For generation of monochromatic light uniform irradiation object to be imaged with excite object radiation fluorescence to be imaged excite the unit, be used for collecting the fluorescence of object radiation to be imaged and be converted into the phosphor collection unit of image information and to carry out the data processing unit of analyzing and processing according to the image information of fluorescence conversion;
Described phosphor collection unit comprises that optics converges parts, photoelectric commutator and is close to the micro optical filter of photoelectric commutator input end, described micro optical filter is multi-channel filter, described micro optical filter comprises a plurality of compact arranged optical filtering infinitesimals, described optical filtering infinitesimal is matrix to be arranged, and a described optical filtering infinitesimal covers a photoelectric conversion unit of described photoelectric commutator; Described optics converges parts and is arranged on the light path of object fluorescent radiation to be imaged, and described micro optical filter is arranged on the reflected light path that described optics converges parts, and the output terminal of described photoelectric commutator connects described data processing unit; Wherein, it is concave mirror that described optics converges parts, and described micro optical filter reaches integrated with it photoelectric commutator and is arranged on the focus of described concave mirror;
Describedly excite irradiation that the unit launches on object to be imaged, object to be imaged is stimulated and gives off the fluorescence of specific wavelength, fluorescence is radiated at described optics and converges on the parts, optics converge that parts converge to micro optical filter with fluorescence and the photoelectric commutator that fits tightly with micro optical filter on, micro optical filter allows the fluorescence signal of specific wavelength to pass through, fluorescence signal is radiated on the photoelectric commutator after by micro optical filter, photoelectric commutator obtains corresponding image information according to described fluorescence signal, and sending image information to described data processing unit, data processing unit is demarcated the information that described photoelectric commutator sends, analyze and process and export the result of analyzing and processing; Described data processing unit is Computerized image processing system, the image information that described photoelectric commutator obtains is delivered to demarcation and the processing that Computerized image processing system carries out image by Computer interface circuit, treat imaging object and carry out real-time analysis and quantitatively detect, and the result of analyzing and processing is transported on the graphoscope shows.
2. little multispectral fluorescence reception according to claim 1 and disposal system, it is characterized in that: the photoelectric commutator of described phosphor collection unit is CCD or cmos image sensor, and described micro optical filter is integrated on CCD or the cmos image sensor.
3. little multispectral fluorescence reception according to claim 1 and disposal system, it is characterized in that: the photoelectric commutator of described phosphor collection unit is for to press the compact arranged photomultiplier of array, and micro optical filter is integrated on the photomultiplier of little gust of arrangement.
4. little multispectral fluorescence reception according to claim 1 and disposal system, it is characterized in that: the described unit that excites comprises LASER Light Source and convex reflecting mirror, described convex reflecting mirror is arranged on the emitting light path of LASER Light Source, and described object to be imaged is arranged on the reflected light path of convex reflecting mirror.
5. little multispectral fluorescence reception according to claim 1 and disposal system, it is characterized in that: the described unit that excites comprises arrowband high-power LED light source and exciting light optical filter, is radiated at equably on the object to be imaged behind the emergent light process exciting light optical filter of described arrowband high-power LED light source.
6. little multispectral fluorescence reception according to claim 1 and disposal system, it is characterized in that: the described unit that excites comprises arrowband high-power LED light source, exciting light optical filter and plane mirror, the emergent light of described arrowband high-power LED light source shines described plane mirror through the exciting light optical filter, and described object to be imaged is positioned on the reflected light path of described plane mirror.
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| CN107525789A (en) * | 2016-06-22 | 2017-12-29 | 株式会社迪思科 | Fluorescence detection device |
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| CN107576263A (en) * | 2017-03-23 | 2018-01-12 | 四川精视科技有限公司 | A kind of vision measurement specular light disturbance restraining method |
| CN108414487B (en) * | 2018-03-08 | 2021-02-12 | 北京航空航天大学 | Optical collection method and system for solid emergent fluorescence |
| CN111272715A (en) * | 2018-12-04 | 2020-06-12 | 长光华大基因测序设备（长春）有限公司 | Fluorescence imaging system of gene sequencer |
| CN111487037A (en) * | 2020-04-16 | 2020-08-04 | 中国科学院微电子研究所 | System and method for detecting uniformity of light source |
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