CN106680186A - Multi-type scattered light detection system of flow cytometer - Google Patents
Multi-type scattered light detection system of flow cytometer Download PDFInfo
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Abstract
The invention discloses a multi-type scattered light detection system of a flow cytometer. The multi-type scattered light detection system comprises a laser and a detection system, wherein the detection system comprises a main light path and a branch light path; detection of the main light path is forward detection; detection of the branch light path is lateral detection; the forward detection and the lateral detection are used for detecting one or more of Rayleigh scattering light, Brillouin scattering light, Raman scattering light, Thomsom scattering light and Compton scattering light; the detection types of the forward detection and the lateral detection can be optionally combined and are not limited by the quantity or the types; the detection directions of the forward detection and the lateral detection can also be regulated in order to adapt to different detection light types. According to the multi-type scattered light detection system of the flow cytometer, disclosed by the invention, different types of scattering light can be detected by utilizing the characteristics of biological samples reflected by the scattering light, more-parameter measurement can carried out on the biological sample, and the measurable range of the biological sample is expanded.
Description
Technical field
The present invention relates to be based on the instrument field of flow cytometer, and in particular to a kind of flow cytometer polymorphic type scattered light
Detection system.
Background technology
Through the development of many decades, current flow cytometer has been realized in commercialization, structure constantly improve, function also day
Benefit is powerful.Fig. 1 show existing flow cytometer schematic diagram.As shown in figure 1, flow cytometer is mainly by four part groups
Into, i.e. flow chamber and liquid fluid system 101, LASER Light Source 102 and optical system 103, Photoelectric Detection and signal processing system 104,
Some flow cytometers further provided with cell sorting system 105.It is now recognized that the flow cytometer structure composition of complete concept should
Including：Flow cytometer main frame, flow cytometer work station, power supply box and automatic sampler (MCL).The main frame of flow cytometer
Structure can be divided into flow chamber, liquid stream drive system, optical system, and signal detection is electronic system with storage, display, analysis system
With five parts such as cell sorting system.
For biological cell, it is different that it is mainly the refractive indexs such as nucleus, Cytoplasm and surrounding medium to scattering of light
Medium refraction, light the difference optical boundary such as cell edges reflection, and the optical diffraction of cell nuclear periphery, intracellular thing
The factors such as the absorption of confrontation light cause.So, can learn You Guan thin by detection and the scattered light signal of analysis biological cell
The information such as born of the same parents' size, shape, internal structure and content content.
In terms of scattered light analysis, scattering light characteristic can be used for the resolution to some cell subgroups.1975
G.C.Salzman etc. is analyzed by measuring the intensity properties of forward scattering light all angles to the backscatter mode of microsphere, and
The forward scattering light intensity for pointing out spherical particle is directly proportional to the volume of ball, and nineteen eighty-three Koichi Shimizu gives scattering
The modification method of intragranular electromagnetic field propagation constant, P.M.A Sloot in 1986 et al. point out that refractive index can be as scattering be to seeing
The distance of measuring point and change, Anastasiya I in 2012 et al. process backscatter mode and realize to all kinds of blood using deconvolution algorithm
The identification of cell.Domestic Jiangsu University Wang Yawei professors team has carried out comprehensive research point to the light scattering characteristic of biological cell
Analysis, such as erythrocyte photon diffusion models, mononuclear cell scattering propertiess, the analysis of complete blood cell phase property, the inspection of biological cell form
Survey etc..University Of Tianjin's Feng Yuan penetrating judgments are awarded and record the dispersion image related to cellular morphology information using CCD, are realized thin to lymph B
Born of the same parents and the identification of T cell, and develop to scattering imaging type flow cytometer.
Sample liquid flow from bottom to top into illumination region when start produce scattered light signal, optical signal is through photodetection system
It is changed into the signal of telecommunication after system, progressively strengthens as the ratio of particle entering part increases current signal, when it progressively flows out illumination
Current signal will progressively reduce until disappearance, during single sample granule flows through illumination region one can be produced again during region
The series of pulses signal of telecommunication.And for different size of cell, under conditions of identical incident illumination and flow velocity, the scattered light of gained
Intensity is simultaneously different, therefore can draw the important letter such as the size of cell by measuring the situation of change of fluidic cell forward scattering light
Breath.
The content of the invention
The invention provides a kind of flow cytometer polymorphic type scattering optical detection system can be in flow cytometer not
The scattered light of same type is detected, and the measurement of flow cytometer multiparameter, energy are realized in the measurement to different types of scattered light
Enough effectively improve flow cytometer systems capacity usage ratio and measurement precision analysis.
The present invention realizes the spy of different types of forward scattering light by changing the spectroscopical number of main optical path front detection
Survey.Forward detection spectroscope is set to n, and n is the positive integer more than or equal to 1, and the Zhi Guanglu of corresponding lateral detection is n+1
Individual, the scattered light light intensity of every Zhi Guanglu is different different with optical path distance, realizes different type side scattered light detections；Often
Bar Zhi Guanglu arranges j spectroscope, and j is the positive integer more than or equal to 1, j+1 scattering optical detection system of the setting for matching, and dissipates
The distance for penetrating photo-detector is different with angle, scatters light to the light intensity difference up to detector, realizes different types of lateral scattered
Penetrate the detection of light.
The technical scheme is that：A kind of flow cytometer polymorphic type scatters optical detection system, including laser instrument and spy
Examining system, it is characterised in that described detection system includes main optical path and Zhi Guanglu；
The main optical path includes n spectroscope and 1 detector for scattered light；
The Zhi Guanglu includes n+1 lens, and described 1 lens, 1 Zhi Guanglu of correspondence, every Zhi Guanglu is provided with j
Spectroscope and j+1 detector for scattered light；
N described spectroscope is sequentially placed into main optical path front, and n+1 lens are respectively placed in n spectroscopical reflection side
To the side with testing sample；
J+1 described scattering optical detection system is respectively placed in j spectroscopical reflection direction with correspondence Zhi Guanglu just
Front；
Each described detector for scattered light is provided with 1 concave lens；Positive integers of the n and j respectively more than or equal to 1.
The laser beam angular that the main optical path detector for scattered light sends with laser instrument is 0 °, and main optical path realizes forward scattering
Optical detection；The Zhi Guanglu detector for scattered light is parallel with the laser beam that laser instrument sends or vertical, and Zhi Guanglu realizes lateral dissipating
Penetrate optical detection.
Preferably, n be 1, j be 4.
Preferably, described concavees lens and the opposing parallel placement of scattering optical detection system；Reflecting mirror and spectroscope and laser
Laser beam out of plumb that device sends and not parallel.
Preferably, the lens be convex lenss, the spectroscope be directly sequentially placed into the rear of lens or spectroscope with it is saturating
Also include reflecting mirror between mirror.
Preferably, detection system detection forward scattering light and side scattered light type are Rayleigh scattering light, Brillouin scattering
Light, Raman diffused light, Thomson scattering light or Compton scattering light.
Preferably, the forward scattering light type of detection system detection is more than one；The side scattered light of detection system detection
Type is more than one.
Preferably, the forward scattering light and side scattered light Type-Inconsistencies of detection system detection.
The invention has the beneficial effects as follows：A kind of flow cytometer polymorphic type scattering optical detection system is dissipated using different types of
Penetrate light to detect the characteristic of biological sample, the measurement of more parameters can be carried out to biological sample, expand tested biological sample
The scope of product.
Description of the drawings
With reference to the accompanying drawing enclosed, the present invention more purpose, function and advantages are by by the as follows of embodiment of the present invention
Description is illustrated, wherein：
Fig. 1 illustrates the structural representation of flow cytometer in prior art；
Fig. 2 illustrates that a kind of flow cytometer polymorphic type of the invention scatters the structural representation of optical detection system；
Fig. 3 illustrates a kind of structural representation of flow cytometer polymorphic type scattering optical detection system embodiment 2 of the invention.
Specific embodiment
By reference to one exemplary embodiment, the purpose of the present invention and function and the side for realizing these purposes and function
Method will be illustrated.However, the present invention is not limited to one exemplary embodiment disclosed below；Can by multi-form come
It is realized.The essence of description is only to aid in the detail of the various equivalent modifications Integrated Understanding present invention.
Hereinafter, embodiments of the invention will be described with reference to the drawings.In the accompanying drawings, identical reference represents identical
Or similar part, or same or like step.
A kind of being achieved in that for flow cytometer polymorphic type scattering optical detection system of the present invention illuminate in flow cytometer
Detector for scattered light is set around hot spot, different directions, the detection of different types of scattered light is realized.
The present invention realizes the spy of different types of forward scattering light by changing the spectroscopical number of main optical path front detection
Survey.Forward detection spectroscope is set to n, and n is the positive integer more than or equal to 1, and the Zhi Guanglu of corresponding lateral detection is n+1
Individual, the scattered light light intensity of every Zhi Guanglu is different different with optical path distance, realizes different type side scattered light detections；Often
Bar Zhi Guanglu arranges j spectroscope, and j is the positive integer more than or equal to 1, j+1 scattering optical detection system of the setting for matching, and dissipates
The distance for penetrating photo-detector is different with angle, scatters light to the light intensity difference up to detector, realizes different types of lateral scattered
Penetrate the detection of light.Due to the spectroscopical setting of forward detection, it is possible to achieve the lateral detection of different branch, lateral detection is realized visiting
Polymorphic type scattered light is surveyed, therefore the present invention can realize the detection of polymorphic type fluidic cell.
Forward detection device detects the size of fluidic cell, and side minder detection fluidic cell granule, forward detection device is visited
The scattered light of survey is the one of Rayleigh scattering light, Brillouin scattering, Raman diffused light, Thomson scattering light or Compton scattering light
Kind, because the parameter that forward detection device and side minder detect fluidic cell is different, therefore forward detection device and laterally detect
Device detection scattered light type is different.Side minder detects the scattered light type in addition to the scattered light of forward detection.
Front side detection detection spectroscope is set to n, and n is the positive integer more than or equal to 1, and n is different so that forward scattering light
The time for reaching forward detection device is different with light intensity, realizes different types of forward scattering optical detection.
Lateral detection is divided into n+1 bar Zhi Guanglu, and each Zhi Guanglu arranges j spectroscope, and j is just whole more than or equal to 1
Number, j+1 scattering optical detection system of the setting for matching, the distance of detector for scattered light is different with angle, scatters light to up to spy
The light intensity for surveying device is different, realizes the detection of different types of side scattered light.
Embodiment 1
Fig. 2 is the structural representation that a kind of flow cytometer polymorphic type of the invention scatters optical detection system.As shown in Figure 2 one
Planting flow cytometer polymorphic type scattering optical detection system includes laser instrument 201, tested biological sample 202 and detection system 203.
Main optical path spectroscope n is 1 in the present embodiment, and the detector for scattered light that moves ahead is 1；Lateral detection Zhi Guanglu is 2,
The spectroscope j of every Zhi Guanglu is 4, and every Zhi Guanglu arranges 5 detector for scattered light.
The main optical path of detection system 203 as shown in Figure 2 includes 1 spectroscope 210 and 1 detector for scattered light 240；Zhi Guang
Road includes 2 lens 220, and every Zhi Guanglu is provided with 4 spectroscopes 230 and 5 detector for scattered light 240.
Spectroscope 210 is placed in main optical path front, and 2 lens 220 are respectively placed in the reflection direction of spectroscope 210 and treat test sample
The side of product 202；
5 scattering optical detection systems 240 be respectively placed in the reflection direction of 4 spectroscopes 230 and correspondence Zhi Guanglu just before
Side；In first spectroscope 230 and it is placed between the lens 220 of the side of testing sample 202 and is provided with a reflecting mirror in Zhi Guanglu
260, for changing light path, it is to avoid produce interference between two Zhi Guanglu detections.
The corresponding opposing parallel placement of scattering optical detection system of concavees lens 250 that each detector for scattered light sets.
Testing sample 202 is placed in the underface of laser instrument 201, and spectroscope 210 is placed in the underface of testing sample, and forward direction dissipates
Penetrate photo-detector and be placed in the underface of spectroscope 210 so the laser beam that sends of main optical path forward scattering photo-detector and laser instrument
Angle is 0 °.The scattering optical detection system of Zhi Guanglu is acted on due to the spectroscope of Zhi Guanglu so that Zhi Guanglu side scattered lights are visited
Survey device parallel with the laser beam that laser instrument sends or vertical.
Wherein, the laser beam that laser instrument 201 sends reaches testing sample 202, and testing sample sends scattered light.Partial dispersion
Light is thrown away and reaches spectroscope 210 along original paths, and the detection of this some scattered light is forward scattering optical detection；Wherein, part
Scattered light Jing spectroscope 210 is transmitted to up to the concavees lens 250 for being placed in the dead ahead of main optical path detector for scattered light 240 and is converged,
Then the light beam after converging is scattered photo-detector and is detected, and another part light is reflected into Zhi Guang through reflecting mirror 210
Road, carries out lateral scattering optical detection.
Another part scattered light is placed in the lateral lens 220 of testing sample 202 and converges, and reflexes to through reflecting mirror 260
Up to first spectroscope 230, which part scattered light is reached to reach and is placed in Zhi Guanglu scattering optical detections through the transmission of spectroscope 230
The concavees lens 250 of the dead ahead of device 240 are converged, and the light beam after then converging is scattered photo-detector 240 and is detected, separately
A part of scattered light is reflexed to up to second spectroscope through first spectroscope 230, wherein second spectroscope is equally to dissipating
Penetrate light to be transmitted and reflected, be a difference in that with the process of first spectroscope 230, scattering light to for Jing transmissions reaches the next one
Spectroscope, reflected scattered light is scattered optical detection system absorption and is detected, and last spectroscope is reached by that analogy,
Reach last spectroscopical scattered light to scatter light to up to the concavees lens for being placed in optical line terminal through what spectroscope was transmitted, pass through
The convergence of concavees lens, is placed in optical line terminal and is mutually perpendicular to the detector for scattered light of placement with other scattered lights of its same branch road
Absorb and detected.
All of detector for scattered light completes light beam detection, and through the process of signal processing system electricity is converted light signals into
Signal, calculator display organization shows the information of simultaneously storage assembly processing system, is further analyzed using flow cytometric.
The detector for scattered light of the forward scattering optical detection is with the detector for scattered light type of lateral scattering optical detection not
Together, the sensitivity of forward scattering photo-detector is less than lateral scattering photo-detector, forward scattering photo-detector reflection fluidic cell
Size, side scattered light reflection fluidic cell granule, therefore detect forward scattering light and side scattered light Type-Inconsistencies.
Forward detection and the scattered light type of lateral detection are Rayleigh scattering light, Brillouin scattering, Raman diffused light, soup
Nurse grandson scattered light or Compton scattering light.
Different types of detector for scattered light is selected in the lateral scattering optical detection, because detector for scattered light is away from walk-off angle
Degree is different so as to which scattered light intensity is different, and the time for reaching detector for scattered light is different, and because different type scatters optical detection
Device selectivity is different so that the side scattered light type of detection system detection is different.Because forward scattering light is stronger, forward direction dissipates
Penetrate reflection and transmission of the light through reflecting mirror so that a small amount of forward scattering light enters detector for scattered light, reaches due to scattering light to
The scattered light intensity of forward detection device is different so that the forward scattering light type of detection system detection is different.
Embodiment 2
Fig. 3 illustrates a kind of structural representation of flow cytometer polymorphic type scattering optical detection system embodiment 2 of the invention.Such as
Shown in Fig. 3, the present embodiment difference compared with embodiment is：The number of the transmitting mirror 310 in main optical path 303 has increased
Plus, the spectroscopical number at Zhi Guanglu 304 has been reduced；It is not provided between lens 340 and first spectroscope anti-
Penetrate mirror.Main optical path spectroscope n is 2 in the present embodiment, and the detector for scattered light that moves ahead is 1；Lateral detection Zhi Guanglu is 3, often
The spectroscope j of bar Zhi Guanglu is 3, and every Zhi Guanglu arranges 4 detector for scattered light.
The present invention realizes the lateral detection of different branch by arranging spectroscope in the detection main optical path that moves ahead；Laterally visiting
Survey branch road and multiple spectroscopes are set, the angle and distance that each spectroscope is arranged is different so that scatter light to up to scattered light and visit
The time of examining system is different with light intensity, and optical detection is scattered to lateral polymorphic type before realizing.
With reference to the explanation of the invention and practice that disclose here, the other embodiment of the present invention is for those skilled in the art
All will be readily apparent and understand.Illustrate and embodiment be to be considered only as it is exemplary, the present invention true scope and purport it is equal
It is defined in the claims.
Claims (7)
1. a kind of flow cytometer polymorphic type scatters optical detection system, including laser instrument and detection system, it is characterised in that described
Detection system include main optical path and Zhi Guanglu；
The main optical path includes n spectroscope and 1 detector for scattered light；
The Zhi Guanglu includes n+1 lens, and described 1 lens, 1 Zhi Guanglu of correspondence, every Zhi Guanglu is provided with j light splitting
Mirror and j+1 detector for scattered light；
N described spectroscope is sequentially placed into main optical path front, n+1 lens be respectively placed in n spectroscopical reflection direction with
The side of testing sample；
J+1 described scattering optical detection system is respectively placed in the dead ahead of j spectroscopical reflection direction and correspondence Zhi Guanglu；
Each described detector for scattered light is provided with 1 concave lens；Positive integers of the n and j respectively more than or equal to 1.
The laser beam angular that the main optical path detector for scattered light sends with laser instrument is 0 °, and main optical path carries out forward scattering light spy
Survey；The detector for scattered light of the Zhi Guanglu is parallel with the laser beam that laser instrument sends or vertical, and the Zhi Guanglu is carried out laterally
Scattering optical detection.
2. flow cytometer polymorphic type according to claim 1 scatters optical detection system, it is characterised in that n is preferably 1, j
Preferably 4.
3. flow cytometer polymorphic type according to claim 1 scatters optical detection system, it is characterised in that described is recessed
Mirror and the opposing parallel placement of scattering optical detection system；Laser beam out of plumb that the spectroscope sends with laser instrument and not parallel.
4. flow cytometer polymorphic type according to claim 1 scatters optical detection system, it is characterised in that the lens are
Convex lenss, the spectroscope of the Zhi Guanglu is directly sequentially placed between the rear of lens or spectroscope and lens and also include reflection
Mirror.
5. flow cytometer polymorphic type according to claim 1 scatters optical detection system, it is characterised in that：Detection system is visited
It is Rayleigh scattering light, Brillouin scattering, Raman diffused light, Thomson scattering light to survey forward scattering light and side scattered light type
Or Compton scattering light.
6. flow cytometer polymorphic type according to claim 5 scatters optical detection system, it is characterised in that：Detection system is visited
The forward scattering light type of survey is more than one, and the side scattered light type of detection system detection is more than one.
7. flow cytometer polymorphic type according to claim 5 scatters optical detection system, it is characterised in that：Detection before to dissipate
Penetrate light and side scattered light Type-Inconsistencies.
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| CN201610951873 | 2016-11-02 | ||
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109556997A (en) * | 2017-09-26 | 2019-04-02 | Ravr有限公司 | The triple channel emerging system of real-time detection particle size and property |
| CN110530783A (en) * | 2018-05-24 | 2019-12-03 | 深圳市帝迈生物技术有限公司 | Lateral light beam collection method, device and flow cytometer for flow cytometer |
| CN115598024A (en) * | 2022-11-16 | 2023-01-13 | 长春光吉科技有限责任公司（Cn） | Cell grading counting device and method thereof |
| US11898951B2 (en) | 2019-08-22 | 2024-02-13 | Hefei Boe Optoelectronics Technology Co., Ltd. | Forward scattered light detection system, flow cytometer and method for measuring cell diameter |
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| US4500641A (en) * | 1981-03-23 | 1985-02-19 | Nederlandse Centrale Organisatie Voor Toegepast Natuurwetenschappelijk Onderzoek | Flow cytometer for identifying algae by chlorophyll fluorescence |
| CN1823269A (en) * | 2003-07-18 | 2006-08-23 | 卢米尼克斯股份有限公司 | Method and system for distinguishing between materials having overlapping spectra |
| CN102998239A (en) * | 2012-06-14 | 2013-03-27 | 龚维燕 | Flow cytometry device for no-flow type cytometry box and flow cytometry method thereof |
| CN103048258A (en) * | 2012-12-26 | 2013-04-17 | 江西科技师范大学 | Light splitting device for flow cytometer |
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|---|---|---|---|---|
| US4500641A (en) * | 1981-03-23 | 1985-02-19 | Nederlandse Centrale Organisatie Voor Toegepast Natuurwetenschappelijk Onderzoek | Flow cytometer for identifying algae by chlorophyll fluorescence |
| CN1823269A (en) * | 2003-07-18 | 2006-08-23 | 卢米尼克斯股份有限公司 | Method and system for distinguishing between materials having overlapping spectra |
| CN102998239A (en) * | 2012-06-14 | 2013-03-27 | 龚维燕 | Flow cytometry device for no-flow type cytometry box and flow cytometry method thereof |
| CN103048258A (en) * | 2012-12-26 | 2013-04-17 | 江西科技师范大学 | Light splitting device for flow cytometer |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109556997A (en) * | 2017-09-26 | 2019-04-02 | Ravr有限公司 | The triple channel emerging system of real-time detection particle size and property |
| CN110530783A (en) * | 2018-05-24 | 2019-12-03 | 深圳市帝迈生物技术有限公司 | Lateral light beam collection method, device and flow cytometer for flow cytometer |
| CN110530783B (en) * | 2018-05-24 | 2023-12-15 | 深圳市帝迈生物技术有限公司 | Lateral beam collection method and device for flow cytometer and flow cytometer |
| US11898951B2 (en) | 2019-08-22 | 2024-02-13 | Hefei Boe Optoelectronics Technology Co., Ltd. | Forward scattered light detection system, flow cytometer and method for measuring cell diameter |
| CN115598024A (en) * | 2022-11-16 | 2023-01-13 | 长春光吉科技有限责任公司（Cn） | Cell grading counting device and method thereof |
| CN115598024B (en) * | 2022-11-16 | 2023-03-14 | 长春光吉科技有限责任公司 | Cell grading counting device and method thereof |
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| CN106680186B (en) | 2019-03-01 |
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