CN103389261A - Spatial parallel type flow cytometry measurement instrument and method - Google Patents
Spatial parallel type flow cytometry measurement instrument and method Download PDFInfo
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- CN103389261A CN103389261A CN2013103123850A CN201310312385A CN103389261A CN 103389261 A CN103389261 A CN 103389261A CN 2013103123850 A CN2013103123850 A CN 2013103123850A CN 201310312385 A CN201310312385 A CN 201310312385A CN 103389261 A CN103389261 A CN 103389261A
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Abstract
The invention discloses a spatial parallel type flow cytometry measurement instrument comprising a sheath flow system, a light source and a polarized light polarization system, wherein n bundles of incident polarized lights with different polarization states are obtained by the polarized light polarization system; the n bundles of incident polarized lights irradiate a sample flow passing through a measurement channel at different positions; different bundles of emergent polarized lights after passing through the measurement channel are received by a polarized light detection system; each bundle of emergent polarized light is subjected to deviation detection by n different polarizers of a deviation detection unit in a corresponding group after being split by a light splitter; n polarization components of each bundle of emergent polarized light are respectively measured; a photoelectric converter converts n*n polarization components obtained by deviation detection into corresponding electric signals; a collecting and processing unit collects and processes the electric signals to generate n*n matrix elements corresponding to the n*n polarization components; n is greater than or equal to 2. The invention also provides a flow cytometry measurement method. By adopting the flow cytometry measurement instrument and method, abundant and accurate microstructure detection information can be obtained.
Description
Technical field
The present invention relates to Flow Cytometry, particularly relate to a kind of fluidic cell measuring instrument and measuring method of spatial parallelism.
Background technology
Flow Cytometry claims again sheath stream technology, is to utilize principle of hydrodynamics to produce sheath stream, and controls sample flow diameter and the flow velocity of sheath stream center, makes the cell that is suspended in sample liquid line up formation along the sample flow center line, successively by the laser acquisition zone.Flow cytometer is automatic cytological analysis and the sorting instrument of a class based on sheath stream technology, it utilizes scattered light and the fluorescent intensity of cell, can obtain size, form, inner structure and specific chemical composition information by the individual cells of search coverage, and accordingly to cell identify, differential count or screening.Flow cytometer is applied in a lot of fields in biological and medical science.Adopt Flow Cytometry to detect one by one Fast Classification to microorganism and become the powerful of microbe research.
Summary of the invention
The object of the invention is to overcome the deficiencies in the prior art, a kind of fluidic cell measuring instrument and measuring method of spatial parallelism is provided, can obtain more horn of plenty and the information of micromechanism detection more accurately.
For achieving the above object, the present invention is by the following technical solutions:
A kind of fluidic cell measuring instrument of spatial parallelism comprises:
The sheath streaming system, be used for making the cell of sample liquid along lining up formation by measuring passage;
Light source and polarized light play inclined to one side system, described polarized light plays inclined to one side system and comprises n different polarizer, each polarizer is polarized the difference bundle in the bundle of the n from light source light separately, to obtain the different n bundle incident polarized light of polarization state, the sample flow that described n bundle incident polarized light passes in different position irradiations the described measurement passage of flowing through;
Polarized light analyzing system, described polarized light analyzing system comprises n group analyzing unit, every group of analyzing unit comprises respectively n different polarizer, each is organized the analyzing unit and is used for receiving the outgoing polarized light of the difference bundle after measuring passage, every bundle outgoing polarized light carries out analyzing by n different polarizer of correspondence group analyzing unit after the optical splitter light splitting, measure respectively n polarized component of every bundle outgoing polarized light;
Photoelectric commutator, be used for and will be converted to corresponding electric signal through n*n the polarized component that analyzing obtains;
The acquisition and processing unit, be used for the described electric signal of acquisition and processing to generate n*n matrix element corresponding to a described n*n polarized component;
N 〉=2 wherein.
Preferably, wherein n is 4,, by obtaining 4*4 matrix element, forms the Muller matrix of 4*4.
Preferably, comprise that being arranged on described polarized light analyzing system is used for the filter plate of the impact of removal fluorescence before.
Preferably, described polarized light plays inclined to one side system and also comprises optical splitter, is used for same light beam is divided into n bundle light, offers respectively n different polarizer.
A kind of fluidic cell measuring method comprises:
Make the cell edge in sample liquid line up the measurement passage of formation by the sheath streaming system;
Use n different polarizer, each polarizer is polarized the difference bundle in the bundle of the n from light source light separately, to obtain the different n bundle incident polarized light of polarization state, the sample flow that described n bundle incident polarized light passes in different position irradiations the described measurement passage of flowing through;
Use n group analyzing unit, every group of analyzing unit comprises respectively n different polarizer, each is organized the analyzing unit and receives the outgoing polarized light of the difference bundle after measuring passage, every bundle outgoing polarized light carries out analyzing through n different polarizer of correspondence group analyzing unit, measures respectively n polarized component of every bundle outgoing polarized light;
Photoelectric commutator will be converted to corresponding electric signal through n*n the polarized component that analyzing obtains;
Acquisition and processing described electric signal in acquisition and processing unit is to generate n*n matrix element corresponding to a described n*n polarized component;
N 〉=2 wherein.
Preferably, wherein n is 4,, by obtaining 4*4 matrix element, forms the Muller matrix of 4*4.
Preferably, removed the impact of fluorescence with filter plate before described polarized light analyzing system.
Preferably, with optical splitter, same light beam is divided into n bundle light and offers respectively n different polarizer.
Useful technique effect of the present invention:
By setting up polarized light to play inclined to one side system and polarized light analyzing system in the fluidic cell measuring instrument, the present invention carries out analyzing after polarized light scatter to the single microbial by detection zone, to be converted to through n*n the polarized component that analyzing obtains corresponding electric signal and final n*n the matrix element that generates corresponding to n*n polarized component, compare traditional scheme, the present invention can get effectively about the more horn of plenty of sample also micromechanism information more accurately.
Description of drawings
Fig. 1 is the structural representation of fluidic cell measuring instrument of the present invention;
Fig. 2 is the structural representation of a kind of specific embodiment of fluidic cell measuring instrument of the present invention.
Embodiment
Below in conjunction with accompanying drawing, embodiments of the invention are elaborated.Should be emphasized that, following explanation is only exemplary, rather than in order to limit the scope of the invention and to apply.
Consult Fig. 1, the fluidic cell measuring instrument comprises that sheath streaming system, light source, polarized light play inclined to one side system, polarized light analyzing system, photoelectric commutator and acquisition and processing unit.Wherein, the sheath streaming system is used for making the cell of sample liquid along lining up formation by measuring passage; Described polarized light plays inclined to one side system and is used for the bundle of the n from light source light is polarized with different polarizers, to obtain the different n bundle incident polarized light of polarization state, described n bundle incident polarization irradiation and the sample flow of passing the described measurement passage of flowing through; Polarized light analyzing system is used for receiving the n bundle outgoing polarized light of described n bundle incident polarized light after by cell scattering, and every bundle outgoing polarized light is carried out analyzing with the individual different polarizer of n, measures respectively n polarized component of every bundle outgoing polarized light; Photoelectric commutator is used for and will be converted to corresponding electric signal through n*n the polarized component that analyzing obtains; The acquisition and processing unit is used for the described electric signal of acquisition and processing to generate n*n matrix element corresponding to a described n*n polarized component.
Consult Fig. 2, wherein, described polarized light plays inclined to one side system and comprises n different polarizer, each polarizer is polarized the difference bundle in the bundle of the n from light source light separately, to obtain the different n bundle incident polarized light of polarization state, the sample flow that described n bundle incident polarized light passes in different position irradiations the described measurement passage of flowing through; Described polarized light analyzing system comprises n group analyzing unit, every group of analyzing unit comprises respectively n different polarizer, each is organized the analyzing unit and is used for receiving the outgoing polarized light of the difference bundle after measuring passage, every bundle outgoing polarized light carries out analyzing by n different polarizer of correspondence group analyzing unit after the optical splitter light splitting, measure respectively n polarized component of every bundle outgoing polarized light.
N 〉=2 wherein, preferably, n is 4, can obtain 4*4 matrix element this moment, thus the Muller matrix of formation 4*4.
In preferred embodiment, described fluidic cell measuring instrument also comprises that being arranged on described polarized light analyzing system is used for removing the filter plate that fluorescence disturbs before.
Other embodiment are about a kind of fluidic cell measuring method, comprise the following steps:
Step S1, make cell in sample liquid along lining up the measurement passage of formation by the sheath streaming system;
Step S2, n different polarizer of use, each polarizer is polarized the difference bundle in the bundle of the n from light source light separately, to obtain the different n bundle incident polarized light of polarization state, the sample flow that described n bundle incident polarized light passes in different position irradiations the described measurement passage of flowing through;
Step S3, use n group analyzing unit, every group of analyzing unit comprises respectively n different polarizer, each is organized the analyzing unit and receives the outgoing polarized light of the difference bundle after measuring passage, every bundle outgoing polarized light carries out analyzing through n different polarizer of correspondence group analyzing unit, measures respectively n polarized component of every bundle outgoing polarized light;
Step S4, photoelectric commutator will be converted to corresponding electric signal through n*n the polarized component that analyzing obtains;
Step S5, the described electric signal of acquisition and processing unit acquisition and processing are to generate n*n matrix element corresponding to a described n*n polarized component;
N 〉=2 wherein.
Preferably, wherein n is 4,, by obtaining 4*4 matrix element, forms the Muller matrix of 4*4.
Preferably, removed the interference of fluorescence with filter plate before light beam is by described polarized light analyzing system.
In the embodiment of the present invention, utilize the sheath streaming system that microbial cell is arranged in team, then the single microbial by detection zone is carried out polarized light scatter.
In preferred embodiment, measure passage for polarized light scatter, the light that light source sends, through beam splitting, rises partially by four different polarization devices respectively, obtains four different polarization states.Four polarization states that one of ordinary skill in the art will readily recognize that inclined to one side four polarization states and analyzing can suitably be chosen according to practical measurement requirement.
Specifically, sample flow in every bundle incident polarization irradiation sheath streaming system and by after single microorganism scattering, with the scattering of α angle and by four different polarizer analyzings, measure respectively four polarized components after corresponding every a branch of incident light scattering, then received and change into electric signal by four photoelectric commutators respectively; 16 road signals are respectively through can, through processing such as amplifications, being gathered by data collecting card; The processor of computing machine is processed the electrical signal data of 16 different incidents of correspondence and scattering polarization state, such as methods such as matrixings, can obtain 16 Muller matrix units, it is as the abundant and Useful Information of analyzing tested cell various features, can be stored and be shown, can also need to make up matrix element, process according to detecting, obtain abundanter detection information and testing result.
Can certainly adopt the matrix of non-4*4., such as only being concerned about the linear polarization part, can only adopt 3 tunnels to rise partially and 3 tunnel analyzings.
Below rise take 4 tunnels partially and 4 tunnel analyzings as example, describe by embodiment more specifically.
as shown in Figure 2, in the present embodiment, described polarized light plays inclined to one side system and comprises 4 different polarizer P1, P2, P3, P4, each polarizer is polarized the difference bundle in 4 bundle light from light source separately, to obtain 4 different bundle incident polarized lights of polarization state, the sample flow that described 4 bundle incident polarized lights pass in different position irradiations the described measurement passage of flowing through, that side of polarized light analyzing comprises 4 groups of analyzings and converting unit D1, D2, D3, D4, every group of analyzing and converting unit comprise respectively 4 different polarizer Ai1, Ai2, Ai3, Ai4(i=1～4), each is organized analyzing and converting unit and is used for receiving the outgoing polarized light of the difference bundle after measuring passage, every bundle outgoing polarized light is first through optical splitter WSout, i(i=1～4) be divided into 4 bundles, respectively by 4 of the analyzing of correspondence group and converting unit different polarizer Ai1, Ai2, Ai3, Ai4(i=1～4) carry out analyzing, measure respectively n polarized component of every bundle outgoing polarized light, again through photodetection module PDi1, PDi2, PDi3, PDi4(i=1～4), by the photomultiplier in the photodetection module, the processing such as pulse amplifier and integrating amplification circuit, obtain electric signal.Then, through analog to digital converter ADC conversion, then deliver to computing machine and process, finally obtain 16 Muller matrix units.
N bundle light from light source can be provided by a plurality of light sources.Can be also to provide light beam by first same light source, described polarized light plays inclined to one side system and can also comprise optical splitter WSin this moment, and this optical splitter WS is divided into 4 bundle light with same light beam, offers respectively 4 different polarizer P1, P2, P3, P4.
, according to the spatial parallelism scheme, be that scattered light, respectively by being received by photoelectric commutator after four different analyzers, obtains four components of scatter light polarization state with the parallel diverse location that is radiated at sample flow in the sheath streaming system of different polarization states incident light.
Can adopt the impulsive measurement circuit, thereby greatly accelerate measurement in single cell speed; And the volume and weight of whole device reduces greatly.In addition, while adopting this scheme, cell carries out time measuring at 4 diverse locations, and likely inverting causes certain error to Muller matrix to be in the variation of spatial orientation of diverse location cell.More preferably, by the sheath current control so that cell is consistent in diverse location time space orientation, thereby reduce error.
Above content is in conjunction with concrete preferred implementation further description made for the present invention, can not assert that specific embodiment of the invention is confined to these explanations.For the general technical staff of the technical field of the invention, without departing from the inventive concept of the premise, can also make some simple deduction or replace, all should be considered as belonging to protection scope of the present invention.
Claims (8)
1. the fluidic cell measuring instrument of a spatial parallelism, is characterized in that, comprising:
The sheath streaming system, be used for making the cell of sample liquid along lining up formation by measuring passage;
Light source and polarized light play inclined to one side system, described polarized light plays inclined to one side system and comprises n different polarizer, each polarizer is polarized the difference bundle in the bundle of the n from light source light separately, to obtain the different n bundle incident polarized light of polarization state, the sample flow that described n bundle incident polarized light passes in different position irradiations the described measurement passage of flowing through;
Polarized light analyzing system, described polarized light analyzing system comprises n group analyzing unit, every group of analyzing unit comprises respectively n different polarizer, each is organized the analyzing unit and is used for receiving the outgoing polarized light of the difference bundle after measuring passage, every bundle outgoing polarized light carries out analyzing by n different polarizer of correspondence group analyzing unit after the optical splitter light splitting, measure respectively n polarized component of every bundle outgoing polarized light;
Photoelectric commutator, be used for and will be converted to corresponding electric signal through n*n the polarized component that analyzing obtains;
The acquisition and processing unit, be used for the described electric signal of acquisition and processing to generate n*n matrix element corresponding to a described n*n polarized component;
N 〉=2 wherein.
2. fluidic cell measuring instrument as claimed in claim 1, is characterized in that, wherein n is 4,, by obtaining 4*4 matrix element, forms the Muller matrix of 4*4.
3. fluidic cell measuring instrument as claimed in claim 1, is characterized in that, comprises that being arranged on described polarized light analyzing system is used for the filter plate of the impact of removal fluorescence before.
4. fluidic cell measuring instrument as described in the claims 1 to 3 any one, is characterized in that, described polarized light plays inclined to one side system and also comprises optical splitter, is used for same light beam is divided into n bundle light, offers respectively n different polarizer.
5. a fluidic cell measuring method, is characterized in that, comprising:
Make the cell edge in sample liquid line up the measurement passage of formation by the sheath streaming system;
Use n different polarizer, each polarizer is polarized the difference bundle in the bundle of the n from light source light separately, to obtain the different n bundle incident polarized light of polarization state, the sample flow that described n bundle incident polarized light passes in different position irradiations the described measurement passage of flowing through;
Use n group analyzing unit, every group of analyzing unit comprises respectively n different polarizer, each is organized the analyzing unit and receives the outgoing polarized light of the difference bundle after measuring passage, every bundle outgoing polarized light carries out analyzing through n different polarizer of correspondence group analyzing unit, measures respectively n polarized component of every bundle outgoing polarized light;
Photoelectric commutator will be converted to corresponding electric signal through n*n the polarized component that analyzing obtains;
Acquisition and processing described electric signal in acquisition and processing unit is to generate n*n matrix element corresponding to a described n*n polarized component;
N 〉=2 wherein.
6. fluidic cell measuring method as claimed in claim 5, is characterized in that, wherein n is 4,, by obtaining 4*4 matrix element, forms the Muller matrix of 4*4.
7. fluidic cell measuring method as claimed in claim 5, is characterized in that, removed the impact of fluorescence before described polarized light analyzing system with filter plate.
8. fluidic cell measuring method as described in claim 5 to 7 any one, is characterized in that, with optical splitter, same light beam is divided into n bundle light and offers respectively n different polarizer.
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| CN106547099A (en) * | 2016-11-02 | 2017-03-29 | 北京信息科技大学 | A kind of flow cytometer beam shaping system based on polarized light |
| CN108681510A (en) * | 2018-05-21 | 2018-10-19 | 迈克医疗电子有限公司 | Data processing method and device |
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| US20050264810A1 (en) * | 2004-04-08 | 2005-12-01 | Tokyo Electron Limited | Particle monitoring device and processing apparatus including same |
| WO2009098867A1 (en) * | 2008-02-07 | 2009-08-13 | Mitsui Engineering & Shipbuilding Co., Ltd. | Fluorescent light detection device and fluorescent light detection method |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106547099A (en) * | 2016-11-02 | 2017-03-29 | 北京信息科技大学 | A kind of flow cytometer beam shaping system based on polarized light |
| CN108681510A (en) * | 2018-05-21 | 2018-10-19 | 迈克医疗电子有限公司 | Data processing method and device |
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