CN103278440A - Cell scattered light multi-position detecting method as well as cell analyzer - Google Patents
Cell scattered light multi-position detecting method as well as cell analyzer Download PDFInfo
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- CN103278440A CN103278440A CN2013101767637A CN201310176763A CN103278440A CN 103278440 A CN103278440 A CN 103278440A CN 2013101767637 A CN2013101767637 A CN 2013101767637A CN 201310176763 A CN201310176763 A CN 201310176763A CN 103278440 A CN103278440 A CN 103278440A
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Abstract
The invention discloses a cell scattered light multi-position detecting method as well as a cell analyzer. The method comprises the following steps of: taking 2-4 detecting points on a cell scattered light plane forming 90 degrees with a laser incidence direction, and detecting 90-degree cell scattered light, wherein the distances from the detecting points to a central line of a sheath flow pipe are equal, and are symmetrical about the central line of the sheath flow pipe. The cell analyzer comprises a laser radiation mechanism, a cell sheath flow mechanism, a scattered light collecting mechanism and a signal analyzing and processing mechanism, wherein the scattered light collecting mechanism comprises a forward scattered light detector and 2-4 90-degree scattered light detectors, and the 90-degree scattered light detectors are arranged on a plane at the laser incidence part vertical to the laser incidence direction, have equal distances from the central line of the sheath flow pipe, and are symmetrical about the central line of the sheath flow pipe. According to the invention, a 90-degree scattered light detecting channel is additionally arranged; the light intensity of an obtained 90-degree scattered light signal is high; the SNR (signal to noise ratio) is improved; the detecting accuracy and the detecting stability are improved; the magnification times and the denoising function of parts of the analyzer can be lowered; the cost of the instrument is lowered; and the reliability is improved.
Description
Technical field
The invention belongs to technical field of biological, be specifically related to a kind of cell scattered light multiposition detection method and cytoanalyze.
Background technology
(flow cytometer FCM) is a kind of cell analysis instrument according to light scattering theory to flow cytometer.During each cell of the single arrangement of in laser shines flow chamber, flowing through because the physical characteristics of cell, part light from the cell through different scattered through angles.Wherein, the light intensity of forward direction small angle scattering light can reflect cell volume; The light intensity of large-angle scattered light can reflect the information of nucleus complexity and cell granulations; And the light intensity of side scattered light can the reacting cells film, nuclear membrane, cytoplasmic variation, thereby cell is classified and cell DAN, RNA is analyzed.
Existing flow cytometer is by sheath stream technology cell to be lined up 1 row, pass through the optical detection window of sheath flow device one by one, the laser that laser instrument produces is radiated at the optical detection window, and individual cells is subjected to the laser irradiation successively, scatters forward direction 0 degree, 10 degree scattered lights and 90 degree scattered lights.Settle a cover pick-up unit on each forward direction and the 90 degree directions, photomultiplier is converted to electric signal with scattered light intensity, for Computer Analysis.
The cell scattered light is original just not strong, also will carry out 2 times, 3 times even light splitting more frequently at 90 scattered lights of spending directions, and polarized light, depolarized light, fluorescent exciting etc. are different to be detected to carry out, and each its intensity of light splitting all weakens 50% at least.Have to can't avoid noise to strengthen with the photomultiplier of high-gain for the 90 degree scattered light information that detect cell, this strengthens the cost of flow cytometer, and the accuracy of whole instrument and stability also are difficult to further raising.
Summary of the invention
The objective of the invention is to design a kind of cell scattered light multiposition detection method, in a plurality of positions of each angle of cell scattered light pick-up unit is set, the cell scattered intensity of acquisition doubles.
Another object of the present invention is that the multiposition detection method according to above-mentioned cell scattered light designs a kind of cell scattered light multiposition and detects cytoanalyze, becoming the multiple spot on 90 ° the scattered light plane of cell that pick-up unit is set with laser direction, to obtain the cell scattered light at double, available photomultiplier than low gain, reduce cost, improve accuracy and the stability of instrument integral body.
When laser is radiated on the individual cells, cell is on the plane that becomes 90 ° with laser direction at the scattered light of 90 ° of directions, on this plane, all can detect 90 ° of scattered lights of cell so, and on this plane, be equal to the cell scattered light information of the cell distance place's of equating detection gained.
Cell scattered light multiposition detection method of the present invention is to become with the laser incident direction on 90 ° the plane of scattered light of cell, getting 2～4 check points, detecting 90 ° of scattered lights of cell.
The check point of 90 ° of scattered lights of described 2～4 cells equates with the distance of sheath stream tube hub line.
When used sheath stream tube section is rectangle, be taken at sheath stream tube wall up and down or about be that 2 points of symmetry are check point with sheath stream tube hub line, perhaps be taken at sheath stream tube wall periphery with sheath stream tube hub line in twos symmetrical 4 points be check point.
Cell scattered light multiposition according to the design of the multiposition detection method of the invention described above cell scattered light detects cytoanalyze, comprise laser radiation mechanism, cellular sheath stream mechanism, scattered light collecting mechanism and signal analysis and processing mechanism, cellular sheath stream mechanism lines up cell to be analyzed in sheath flow liquid single-row along its sheath stream pipe streamlined flow, the laser that laser radiation mechanism produces shines successively to the cell in the sheath stream pipe, the laser incident direction is vertical with sheath stream pipe trend, the scattered light collecting mechanism comprises 1 cover forward scattering photodetector and 90 ° of light scattering detectors of 2～4 covers, the forward scattering photodetector be arranged at laser in the same way, 10 ° of positions, the place ahead, 90 ° of light scattering detectors are arranged on the laser incident place plane vertical with the laser incident direction.
Each 90 ° of light scattering detector present position equates with the distance of this sheath stream tube hub line.Can adopt identical light scattering detector module.
When used sheath stream tube section is rectangle, 90 ° of light scattering detectors on the laser incident place plane vertical with the laser incident direction be positioned at sheath stream tube wall up and down or about be 2 points of symmetry with sheath stream tube hub line, perhaps be positioned at sheath stream tube wall periphery with sheath stream tube hub line symmetrical 4 points in twos.
Preferred plan for be positioned at 90 ° of light scattering detectors on the laser incident place plane vertical with the laser incident direction about sheath stream tube wall to flow the tube hub line with sheath be 2 points of symmetry, this line of 2 is horizontal line.
Compared with prior art, the advantage of cell scattered light multiposition detection method of the present invention and cytoanalyze is: 1, on the basis of former detection light path, increased the sense channel of 90 ° of scattered lights, 90 ° of scattered light signal light intensity of acquisition are intrinsic 2～4 times; 2, owing to 90 ° of scattered light signal light intensity height that obtain, signal to noise ratio (S/N ratio) improves, the accuracy of cell analysis and stability raising; 3, need not the photomultiplier of adapted high-gain, reduced the requirement to equipment circuit enlargement factor and noise removal function, instrument cost reduces, and reliability improves.
Description of drawings
Fig. 1 is this cell scattered light multiposition detection method embodiment principle schematic;
Fig. 2 is that this cell scattered light multiposition detects cytoanalyze example structure synoptic diagram.
Number in the figure is:
1, laser radiation mechanism, 2,90 ° of light scattering detectors, 3, cellular sheath stream mechanism, 4, the forward scattering photodetector.
Embodiment
Further describe the present invention below in conjunction with the drawings and specific embodiments.
Cell scattered light multiposition detection method embodiment
As shown in Figure 1, laser is advanced along X-direction, and sheath stream tube hub line is along the Y direction setting, and sheath stream tube section is rectangle, and tube wall is parallel to the XY plane before and after it, and left and right sides tube wall is vertical with the laser direct of travel.When the individual cells that is arranged in rows during through laser, the cell that is subjected to the laser irradiation is at the scattered light of 90 ° of directions on the plane P that becomes 90 ° with laser direction, namely Fig. 1 the YZ plane.
On plane P, be equal to the cell scattered light information of detection gained on the point that sheath stream tube hub linear distance equates.This example is got A, the B, C, the D4 point that equate with sheath stream tube hub linear distance and is check point, detects 90 ° of scattered lights through the true origin cell.A, a B2 check point is on the Z axle, and symmetrical centered by true origin, C, a D2 check point is on plane P, and line and the Y-axis of C, D and initial point are at 45, are rotational symmetry with the Y-axis.
Cell scattered light multiposition detects cytoanalyze embodiment
This is for example shown in Figure 2, this routine cytoanalyze by last routine cell scattered light multiposition detection method design comprises laser radiation mechanism 1, cellular sheath stream mechanism 3, scattered light collecting mechanism and signal analysis and processing mechanism, and the laser that laser radiation mechanism 1 produces is advanced along X-axis; Cellular sheath stream mechanism 3 makes cell to be analyzed line up single-row stream along its sheath in sheath flow liquid and manages streamlined flow, and sheath stream pipe is along the Y direction setting, and sheath stream tube section is rectangle, and tube wall is parallel to the XY plane before and after it, and left and right sides tube wall is vertical with the laser direct of travel.The intersection point of laser and sheath stream pipe is true origin.The laser that laser radiation mechanism 1 produces shines successively to the cell of the process true origin in the sheath stream pipe.
The scattered light collecting mechanism comprises three cover light scattering detectors.Through the 10 ° of positions, the place ahead after the true origin, namely the F of Fig. 2 point is provided with forward scattering photodetector 4 at X-axis laser.At Z-direction, A, B point in about sheath stream pipe 90 ° of light scattering detectors 2 of a cover are set respectively, A, B point and initial point are apart from equating.A, B point is symmetry on the Z axle, with the true origin.The line of this routine AB is horizontal line.
Above-described embodiment is the specific case that purpose of the present invention, technical scheme and beneficial effect are further described only, and the present invention is defined in this.All any modifications of within scope of disclosure of the present invention, making, be equal to replacement, improvement etc., all be included within protection scope of the present invention.
Claims (7)
1. cell scattered light multiposition detection method is becoming with laser direction on 90 ° the plane of scattered light of cell, and getting 2～4 points is check point, detects 90 ° of scattered lights of cell.
2. cell scattered light multiposition detection method according to claim 1 is characterized in that:
The check point of 90 ° of scattered lights of described 2～4 cells equates with the distance of sheath stream tube hub line.
3. cell scattered light multiposition detection method according to claim 2 is characterized in that:
When used sheath stream tube section is rectangle, be taken at sheath stream tube wall up and down or about be that 2 points of symmetry are check point with sheath stream tube hub line, perhaps be taken at sheath stream tube wall periphery with sheath stream tube hub line in twos symmetrical 4 points be check point.
4. the cell scattered light multiposition of cell scattered light multiposition detection method design according to claim 1 detects cytoanalyze, comprise laser radiation mechanism (1), cellular sheath stream mechanism (3), scattered light collecting mechanism and signal analysis and processing mechanism, cellular sheath stream mechanism (3) lines up cell to be analyzed in sheath flow liquid single-row along its sheath stream pipe streamlined flow, the laser that laser radiation mechanism (1) produces shines successively to the cell in the sheath stream pipe, the laser incident direction is vertical with sheath stream pipe trend, the scattered light collecting mechanism comprises 1 cover forward scattering photodetector (4), it is characterized in that:
Also have 2～4 cover 90 ° of light scattering detectors (2), 90 ° of light scattering detectors (2) are arranged on the laser incident place plane vertical with the laser incident direction.
5. cell scattered light multiposition according to claim 4 detects cytoanalyze, it is characterized in that:
Described each 90 ° of light scattering detectors (2) present position equates with the distance of sheath stream tube hub line.
6. cell scattered light multiposition according to claim 5 detects cytoanalyze, it is characterized in that:
Described sheath stream tube section is rectangle, the 90 ° of light scattering detectors (2) on the vertical plane of laser incident place and sheath stream pipe be positioned at sheath stream tube wall up and down or about be 2 points of symmetry with sheath stream tube hub line, perhaps be positioned at sheath stream tube wall periphery with sheath stream tube hub line symmetrical 4 points in twos.
7. cell scattered light multiposition according to claim 5 detects cytoanalyze, it is characterized in that:
Be positioned at 2 symmetric points about sheath stream tube wall at the 90 ° of light scattering detectors (2) on the laser incident place plane vertical with the laser incident direction, the line of 2 symmetric points is horizontal line.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103575637A (en) * | 2013-11-12 | 2014-02-12 | 桂林优利特医疗电子有限公司 | Sheath flow impedance and optical synchronous counting device |
| CN105572017A (en) * | 2015-12-21 | 2016-05-11 | 深圳联开生物医疗科技有限公司 | Smart gain adjustment system and blood cell analyzer |
| CN114047101A (en) * | 2021-07-12 | 2022-02-15 | 中国科学院大气物理研究所 | Optical simulation system and method for representing irregularity degree of particulate matter |
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| CN1116708A (en) * | 1994-08-08 | 1996-02-14 | 东亚医用电子株式会社 | Cells analysing device |
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| EP2333525A1 (en) * | 2008-09-19 | 2011-06-15 | Mitsui Engineering & Shipbuilding Co., Ltd. | Fluorescence detection device by means of intensity moludated laser light and method for detecting fluorscence |
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| CN203249851U (en) * | 2013-05-14 | 2013-10-23 | 桂林优利特医疗电子有限公司 | Cell scattering light multi-position detection cytometer |
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- 2013-05-14 CN CN2013101767637A patent/CN103278440A/en active Pending
Patent Citations (6)
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| CN1116708A (en) * | 1994-08-08 | 1996-02-14 | 东亚医用电子株式会社 | Cells analysing device |
| CN101236158A (en) * | 2007-01-29 | 2008-08-06 | 深圳迈瑞生物医疗电子股份有限公司 | Reticular cell detection method and detection device |
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| EP2333525A1 (en) * | 2008-09-19 | 2011-06-15 | Mitsui Engineering & Shipbuilding Co., Ltd. | Fluorescence detection device by means of intensity moludated laser light and method for detecting fluorscence |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103575637A (en) * | 2013-11-12 | 2014-02-12 | 桂林优利特医疗电子有限公司 | Sheath flow impedance and optical synchronous counting device |
| CN105572017A (en) * | 2015-12-21 | 2016-05-11 | 深圳联开生物医疗科技有限公司 | Smart gain adjustment system and blood cell analyzer |
| CN105572017B (en) * | 2015-12-21 | 2018-07-06 | 深圳联开生物医疗科技有限公司 | A kind of Intelligent Gain regulating system and cellanalyzer |
| CN114047101A (en) * | 2021-07-12 | 2022-02-15 | 中国科学院大气物理研究所 | Optical simulation system and method for representing irregularity degree of particulate matter |
| CN114047101B (en) * | 2021-07-12 | 2022-06-10 | 中国科学院大气物理研究所 | Optical simulation system and method for representing irregularity degree of particulate matter |
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Application publication date: 20130904 |