CN106680186B - A kind of flow cytometer polymorphic type scattering optical detection system - Google Patents
A kind of flow cytometer polymorphic type scattering optical detection system Download PDFInfo
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- 238000001514 detection method Methods 0.000 title claims abstract description 106
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- 238000001069 Raman spectroscopy Methods 0.000 claims abstract description 5
- 239000012472 biological sample Substances 0.000 abstract description 7
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- 210000004027 cell Anatomy 0.000 description 19
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- G—PHYSICS
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
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Abstract
The invention discloses a kind of flow cytometer polymorphic types to scatter optical detection system, which includes laser and detection system, and the detection system includes main optical path and Zhi Guanglu, and main optical path detection is forward detection, and Zhi Guanglu is lateral detection；Forward detection and lateral detection detect one or more of Rayleigh scattering light, Brillouin scattering, Raman diffused light, Thomson scattering light and Compton scattering light, the detection type of the two can be combined arbitrarily, it is not limited by quantity or classification, the detection direction of the two can also be adjusted to adapt to different detection light types.The characteristic for the biological sample that the present invention is reflected using different types of scattering light detects it, and the measurement of more parameters can be carried out to biological sample, expands the range that can survey biological sample.
Description
Technical field
The present invention relates to the instrument fields based on flow cytometer, and in particular to a kind of flow cytometer polymorphic type scattering light
Detection system.
Background technique
By the development of many decades, flow cytometer has been realized in commercialization at present, and 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
At, 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 are further provided with cell sorting system 105.It is now recognized that the flow cytometer structure composition of complete concept is answered
It include: flow cytometer host, flow cytometer work station, power supply box and autosampler (MCL).The host of flow cytometer
Structure can be divided into flow chamber, liquid stream drive system, optical system, signal detection and storage, display, analysis system, that is, electronic system
With five parts such as cell sorting system.
For biological cell, the scattering to light is mainly that the refractive index such as nucleus, cytoplasm and surrounding medium are different
Medium refraction, light is in the different reflection of optical boundary such as cell edges and the optical diffraction of cell nuclear periphery, intracellular object
The factors such as the absorption of confrontation light cause.So can be learnt by the scattered light signal for detecting with analyzing biological cell You Guan thin
The information such as born of the same parents' size, shape, internal structure and content content.
In terms of scattering light analysis, scattering light characteristic can be used for the resolution to certain cell subgroups.1975
G.C.Salzman etc. analyzes the backscatter mode of microballoon by measuring the intensity properties of forward scattering light all angles, and
Point out that the forward scattering luminous intensity of spherical particle is directly proportional to the volume of ball, 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 with scattering to sight
The distance of measuring point and change, Anastasiya I in 2012 et al. using deconvolution algorithm processing backscatter mode realize to all kinds of blood
The identification of cell.Domestic Jiangsu University Wang Yawei professor team has carried out comprehensive research point to the light scattering characteristic of biological cell
Analysis, such as the analysis of red blood cell photon diffusion models, monocyte scattering properties, whole blood cells phase property, the inspection of biological cell form
Survey etc..University Of Tianjin's Feng Yuan penetrating judgment, which is awarded, records dispersion image relevant to cellular morphology information using CCD, realizes thin to lymph B
The identification of born of the same parents and T cell, and scattering imaging type flow cytometer is developed.
Sample liquid flow starts to generate scattered light signal when entering illumination region from bottom to top, and optical signal passes through photodetection system
It is changed into electric signal after system, enhances as the ratio of particle entering part increases current signal gradually, when it gradually flows out illumination
Current signal will be gradually reduced again until disappearance, can generate one during single sample particle flows through illumination region when region
Series of pulses electric signal.And for different size of cell, under conditions of identical incident light and flow velocity, resulting scattering light
Intensity is simultaneously different, therefore can obtain the important letter such as size of cell by the situation of change of measurement fluidic cell forward scattering light
Breath.
Summary of the invention
The present invention provides a kind of flow cytometer polymorphic type scattering optical detection systems can be in flow cytometer not
The scattering light of same type is detected, and the measurement of flow cytometer multi-parameter, energy are realized to the measurement of different types of scattering light
Enough effectively improve flow cytometer systems capacity usage ratio and measurement precision analysis.
The present invention detects spectroscopical number by changing in front of main optical path, realize the spy of different types of forward scattering light
It surveys.Forward detection spectroscope is set as n, and n is the positive integer more than or equal to 1, and the corresponding Zhi Guanglu laterally detected is n+1
A, scattering light luminous intensity difference and the optical path distance difference of every Zhi Guanglu realize different type lateral scattering optical detections；Often
J spectroscope is arranged in Zhi Guanglu, and j is the positive integer more than or equal to 1, and j+1 scattering optical detection system of the setting to match is scattered
The distance and angle difference for penetrating optical detector scatter the luminous intensity difference that light reaches detector, realize 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 and spy
Examining system, which is characterized in that the 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 1 lens corresponding 1 Zhi Guanglu, every Zhi Guanglu are equipped with j
Spectroscope and j+1 detector for scattered light；
The n spectroscope is sequentially placed into front of main optical path, and n+1 lens are respectively placed in n spectroscopical reflection sides
To the side with sample to be tested；
The j+1 scattering optical detection system is being respectively placed in j spectroscopical reflection directions and corresponding Zhi Guanglu just
Front；
Each detector for scattered light is equipped with 1 concave lens；N and j is respectively the positive integer for being more than or equal to 1.
The laser beam angular that the main optical path detector for scattered light and laser issue is 0 °, and main optical path realizes forward scattering
Optical detection；The Zhi Guanglu detector for scattered light is parallel or vertical with the laser beam that laser issues, and Zhi Guanglu realizes lateral dissipate
Penetrate optical detection.
Preferably, 1 n, j 4.
Preferably, optical detection system is opposite is placed in parallel with scattering for the concavees lens；Reflecting mirror and spectroscope and laser
The laser beam out of plumb and not parallel that device issues.
Preferably, the lens be convex lens, the spectroscope be directly sequentially placed into lens rear or spectroscope and thoroughly
It further include reflecting mirror between mirror.
Preferably, detection system detection forward scattering light and lateral scattering 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 beneficial effects of the present invention are: a kind of flow cytometer polymorphic type scattering optical detection system is dissipated using different types of
It penetrates 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 range of product.
Detailed description of the invention
With reference to the attached drawing of accompanying, the more purposes of the present invention, function and advantage are by the as follows of embodiment through the invention
Description is illustrated, in which:
Fig. 1 shows the structural schematic diagram of flow cytometer in the prior art；
Fig. 2 shows a kind of structural schematic diagrams of flow cytometer polymorphic type scattering optical detection system of the present invention；
Fig. 3 shows a kind of structural schematic diagram of flow cytometer polymorphic type scattering optical detection system embodiment 2 of the present invention.
Specific embodiment
By reference to 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 exemplary embodiment as disclosed below；Can by different form come
It is realized.The essence of specification is only to aid in those skilled in the relevant arts' Integrated Understanding detail of the invention.
Hereinafter, the embodiment of the present invention will be described with reference to the drawings.In the accompanying drawings, identical appended drawing reference represents identical
Or similar component 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 illuminates in flow cytometer
Detector for scattered light is set around hot spot, realizes the detection of different directions, different types of scattering light.
The present invention detects spectroscopical number by changing in front of main optical path, realize the spy of different types of forward scattering light
It surveys.Forward detection spectroscope is set as n, and n is the positive integer more than or equal to 1, and the corresponding Zhi Guanglu laterally detected is n+1
A, scattering light luminous intensity difference and the optical path distance difference of every Zhi Guanglu realize different type lateral scattering optical detections；Often
J spectroscope is arranged in Zhi Guanglu, and j is the positive integer more than or equal to 1, and j+1 scattering optical detection system of the setting to match is scattered
The distance and angle difference for penetrating optical detector scatter the luminous intensity difference that light reaches detector, realize different types of lateral scattered
Penetrate the detection of light.Due to the spectroscopical setting of forward detection, the lateral detection of different branch may be implemented, lateral detection, which is realized, to be visited
It surveys polymorphic type and scatters light, therefore the detection of polymorphic type fluidic cell may be implemented in the present invention.
Forward detection device detects the size of fluidic cell, and side minder detects fluidic cell particle, and forward detection device is visited
The scattering light of survey is the one of Rayleigh scattering light, Brillouin scattering, Raman diffused light, Thomson scattering light or Compton scattering light
Kind, since forward detection device is different with the parameter of side minder detection fluidic cell, forward detection device and lateral detection
Device detection scattering light type is different.Side minder detects the scattering light type in addition to the scattering light of forward detection.
Front side detection detection spectroscope is set as n, and n is the positive integer more than or equal to 1, and n is different, so that forward scattering light
Time and the luminous intensity difference for reaching forward detection device, realize different types of forward scattering optical detection.
Lateral detection is divided into n+1 Zhi Guanglu, and j spectroscope is arranged in each Zhi Guanglu, and j is just whole more than or equal to 1
Number, j+1 scattering optical detection system of the setting to match, distance and the angle difference of detector for scattered light, scattering light, which reaches, to be visited
The luminous intensity for surveying device is different, realizes the detection of different types of side scattered light.
Embodiment 1
Fig. 2 is the structural schematic diagram that a kind of flow cytometer polymorphic type of the present invention scatters optical detection system.As shown in Figure 2 one
Kind flow cytometer polymorphic type scattering optical detection system includes laser 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 5 detector for scattered light are arranged in every Zhi Guanglu.
203 main optical path of detection system 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 equipped with 4 spectroscope 230 and 5 detector for scattered light 240.
Spectroscope 210 is placed in front of main optical path, and 2 lens 220 are respectively placed in the reflection direction of spectroscope 210 and to test sample
The side of product 202；
5 scattering optical detection systems 240 be respectively placed in 4 spectroscopes 230 reflection direction and corresponding Zhi Guanglu just before
Side；In Zhi Guanglu in first spectroscope 230 and be placed between the lens 220 of 202 side of sample to be tested be equipped with a reflecting mirror
260, for changing optical path, avoid generating interference between two Zhi Guanglu detections.
The corresponding scattering optical detection system of the concavees lens 250 that each detector for scattered light is set is opposite to be placed in parallel.
Sample to be tested 202 is placed in the underface of laser 201, and spectroscope 210 is placed in the underface of sample to be tested, and forward direction dissipates
It penetrates optical detector and is placed in the underface of spectroscope 210 so the laser beam that main optical path forward scattering optical detector and laser issue
Angle is 0 °.The scattering optical detection system of Zhi Guanglu is since the spectroscope of Zhi Guanglu acts on, so that Zhi Guanglu side scattered light is visited
It is parallel or vertical with the laser beam that laser issues to survey device.
Wherein, the laser beam that laser 201 issues reaches sample to be tested 202, and sample to be tested issues scattering light.Partial dispersion
Light continues to reach spectroscope 210 along original paths, and the detection of this some scattered light is forward scattering optical detection；Wherein, portion
Dispersion is penetrated the concavees lens 250 that light is placed in 240 front of main optical path detector for scattered light through the transmission arrival of spectroscope 210 and is converged
Poly-, the light beam after then converging is detected by detector for scattered light, and another part light is reflected into branch by reflecting mirror 210
Optical path carries out lateral scattering optical detection.
Another part scattering light be placed in 202 side of sample to be tested to lens 220 converge, be reflected by reflecting mirror 260
Up to first spectroscope 230, part scattering light is transmitted to by spectroscope 230 is placed in Zhi Guanglu detector for scattered light 240
The concavees lens 250 in front are converged, and the light beam after then converging is detected by detector for scattered light 240, another portion
Dispersion penetrates light and reaches second spectroscope by first reflection of spectroscope 230, wherein second spectroscope is equally to scattering light
It is transmitted and is reflected, the process with first spectroscope 230 is except that the scattering light through transmiting reaches next light splitting
Mirror, scattering light through reflection by scattering optical detection system absorption is detected, and so on reach the last one spectroscope, reach
The last one the scattering light of spectroscopical scattering light by spectroscope transmission reaches the concavees lens for being placed in optical line terminal, by recessed
The convergence of mirror, other scattering light for being placed in optical line terminal and its same branch are mutually perpendicular to the detector for scattered light placed absorption
And it is detected.
All detector for scattered light complete light beam detection, convert light signals into electricity by the processing of signal processing system
Signal, calculator display organization is shown and the information of storage assembly processing system, is further analyzed using flow cytometric.
The detector for scattered light of the forward scattering optical detection and the detector for scattered light type of lateral scattering optical detection are not
Together, the sensitivity of forward scattering optical detector is less than lateral scattering optical detector, and forward scattering optical detector reflects fluidic cell
Size, side scattered light reflects fluidic cell particle, therefore detects forward scattering light and side scattered light Type-Inconsistencies.
Forward detection and the scattering light type laterally detected are Rayleigh scattering light, Brillouin scattering, Raman diffused light, soup
Nurse grandson scatters light or Compton scattering light.
Different types of detector for scattered light is selected in the lateral scattering optical detection, since detector for scattered light is away from walk-off angle
Degree is different, keeps its scattered light intensity different, and the time for reaching detector for scattered light is different, and since different type scatters optical detection
Device selectivity is different, so that the lateral scattering light type of detection system detection is different.Since forward scattering light is stronger, forward direction dissipates
Reflection and transmission that light passes through reflecting mirror are penetrated, so that a small amount of forward scattering light enters detector for scattered light, since scattering light reaches
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 shows a kind of structural schematic diagram of flow cytometer polymorphic type scattering optical detection system embodiment 2 of the present 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 is increased
Add, spectroscopical number at Zhi Guanglu 304 is 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 Zhi Guanglu is 3, and 4 detector for scattered light are arranged in every Zhi Guanglu.
The present invention realizes the lateral detection of different branch by the way that spectroscope is arranged in the detection main optical path that moves ahead；Laterally visiting
It surveys branch and multiple spectroscopes is set, the angle and distance of each spectroscope setting is different, so that scattering light reaches scattering light and visits
The time of examining system and luminous intensity difference, scatter optical detection to lateral polymorphic type before realizing.
In conjunction with the explanation and practice of the invention disclosed here, the other embodiment of the present invention is for those skilled in the art
It all will be readily apparent and understand.Illustrate and embodiment is regarded only as being exemplary, true scope of the invention and purport are equal
It is defined in the claims.
Claims (7)
1. a kind of flow cytometer polymorphic type scatters optical detection system, including laser and detection system, which is characterized 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, wherein 1 lens corresponding 1 Zhi Guanglu, every Zhi Guanglu are equipped with j spectroscope
With j+1 detector for scattered light；
The n spectroscope is sequentially placed into front of main optical path, n+1 lens be respectively placed in n spectroscopical reflection directions with
The side of sample to be tested；
The j+1 detector for scattered light is respectively placed in the front of j spectroscopical reflection directions and corresponding Zhi Guanglu；
Wherein each detector for scattered light is equipped with 1 concave lens；N and j is respectively the positive integer for being more than or equal to 1；
The laser beam angular that wherein detector for scattered light of main optical path and laser issue is 0 °, and main optical path carries out forward scattering light
Detection；The detector for scattered light of Zhi Guanglu is parallel or vertical with the laser beam that laser issues, and the Zhi Guanglu is laterally dissipated
Penetrate optical detection.
2. flow cytometer polymorphic type according to claim 1 scatters optical detection system, which is characterized in that n 1, j 4.
3. flow cytometer polymorphic type according to claim 1 scatters optical detection system, which is characterized in that described is recessed
Mirror is opposite with detector for scattered light to be placed in parallel；Laser beam out of plumb that the spectroscope and laser issue and not parallel.
4. flow cytometer polymorphic type according to claim 1 scatters optical detection system, which is characterized in that the lens are
Convex lens, it further includes reflection that the spectroscope of the Zhi Guanglu, which is directly sequentially placed between the rear or spectroscope and lens of lens,
Mirror.
5. flow cytometer polymorphic type according to claim 1 scatters optical detection system, it is characterised in that: detection system is visited
It surveys forward scattering light and lateral scattering light type is Rayleigh scattering light, Brillouin scattering, Raman diffused light, Thomson scattering light
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 lateral scattering 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: to scattered before detection
Penetrate light and side scattered light Type-Inconsistencies.
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| 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 |
| 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 |
| CN110411933B (en) | 2019-08-22 | 2022-04-26 | 合肥京东方光电科技有限公司 | Forward scattering light detection system, flow cytometer, and method for measuring cell diameter |
| 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 |
Citations (4)
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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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2017
- 2017-01-19 CN CN201710044726.9A patent/CN106680186B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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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