CN103033464A - Photoacoustic-fluorescence flow cytometer - Google Patents
Photoacoustic-fluorescence flow cytometer Download PDFInfo
- Publication number
- CN103033464A CN103033464A CN2012105741222A CN201210574122A CN103033464A CN 103033464 A CN103033464 A CN 103033464A CN 2012105741222 A CN2012105741222 A CN 2012105741222A CN 201210574122 A CN201210574122 A CN 201210574122A CN 103033464 A CN103033464 A CN 103033464A
- Authority
- CN
- China
- Prior art keywords
- fluorescence
- flow cytometer
- optoacoustic
- signal receiving
- computer display
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Images
Abstract
The invention discloses a photoacoustic-fluorescence flow cytometer. The flow cytometer comprises an optical excitation system, a fluorescence signal receiving system, a photoacoustic signal receiving system, a sample platform and a computer display controlling system, wherein the optical excitation system is connected with the fluorescence signal receiving system, the sample platform and the computer display controlling system; the photoacoustic signal receiving system is connected with the sample platform and the computer display controlling system; the fluorescence signal receiving system is connected with the computer display controlling system; the computer display controlling system is electrically connected with the optical excitation system, the fluorescence signal receiving system and the photoacoustic signal receiving system. According to the flow cytometer, as a pulse laser is adopted as a single excitation light source, a photoacoustic signal and a fluorescence signal of a substance to be detected are excited simultaneously, and simultaneously acquired through an upper passage and a lower passage, the photoacoustic signal and the fluorescence signal are detected in real time, and more information about a single cell and granules is acquired.
Description
Technical field
The invention belongs to the flow cytometer detection technical field, particularly a kind of optoacoustic-fluorescence flow cytometer and detection method and application.
Background technology
Flow cytometry is the technology that the seventies grows up, it integrates computer technology, laser technology, fluid mechanics, cytochemistry, cellular immunology, have simultaneously and analyze and the function of sorting cells, this technology be a kind of on functional level to detection means unicellular or that particle carries out quantitative test and sorting.
At present, what the fluidic cell detection method was the most frequently used is the fluorescence flow cytometry, it shines under the flow at high speed state by the cell of fluorescent dyeing or particle with continuous laser, measure the intensity of its emitting fluorescence, thereby reach a kind of cell analysis technology of cell or particle being carried out qualitative or quantitative determination and analysis.The fluorescence flow cytometer is widely used in the fields such as biomedicine, molecular biology at present.Yet owing to rely on the fluoroscopic examination of extrinsic fluorescence dyestuff, there is certain limitation in its method: (1) can't be detected dark matter (non-fluorescent material); (2) fluorescent dye in cell may with the physiological status of cell change degrade, the phenomenons such as decomposition or cancellation, thereby can't detect fluorescence and undetected.
When optoacoustic effect refers to that short light pulse shines certain absorber, absorbent body luminous energy and produce the moment temperature rise, temperature rise causes the volume breathing of absorber, thereby gives off ultrasound wave.Optoacoustic effect produces and to depend on the absorption coefficient of light of exciting light parameter and irradiation object, as long as have the material of light absorption effectively to be converted into ultrasonic signal and be detected, need not depend on fluorescent material.Such as the non-fluorescent material with the endogenous pigment molecule such as red blood cell, by photo-acoustic excitation, detect the ultrasonic optoacoustic flow cytometer detection of just can realizing that radiates, and need not the again fluorchrome of mark external source.Therefore, the optoacoustic flow cytometer detection is combined with the method for fluorescence flow cytometer detection, utilize same light source to excite simultaneously optoacoustic and fluorescence, and detect simultaneously the signal of two kinds of energy models, just can realize the flow cytometer detection art of optoacoustic-fluorescent dual module attitude, can effectively remedy the deficiency of fluorescence flow cytometer detection method, realize bimodal, multiparameter, complementary flow cytometer detection new technology.
Summary of the invention
The shortcoming that primary and foremost purpose of the present invention is to overcome prior art provides a kind of optoacoustic-fluorescence flow cytometer with not enough.The advantage of this flow cytometer is to use pulse laser as single excitation source, optoacoustic and the fluorescence signal of while excitation-detection material, then pass through up and down two paths, gather simultaneously respectively optoacoustic and the fluorescence signal of detected material, realize two kinds of multi-functional flow cytometer detections of mode.
Another object of the present invention is to provide the detection method of using above-mentioned optoacoustic-fluorescence flow cytometer.
A further object of the present invention is to provide the application of described optoacoustic-fluorescence flow cytometer.
Purpose of the present invention is achieved through the following technical solutions: a kind of optoacoustic-fluorescence flow cytometer comprises optical excitation system, fluorescence signal receiving system, photoacoustic signal receiving system, sample stage and computing machine (PC) display control program; The optical excitation system is connected with fluorescence signal receiving system, sample stage, Computer display control system respectively, the photoacoustic signal receiving system is connected with sample stage, Computer display control system respectively, and the fluorescence signal receiving system is connected with the Computer display control system; The Computer display control system is electrically connected with optical excitation system, fluorescence signal receiving system, photoacoustic signal receiving system respectively;
Described optical excitation system comprises excitation source, diaphragm, cylindrical lens, slit A, achromat A, catoptron, dichroic beam splitter A and microcobjective; Excitation source and the electrical connection of Computer display control system; Diaphragm, cylindrical lens, slit A, achromat A, catoptron, dichroic beam splitter A and microcobjective be mechanical connection successively; Microcobjective and described sample stage mechanical connection; The optical excitation system device all is fixed in the camera bellows, and diaphragm, cylindrical lens, slit A, achromat A all can move in the camera bellows track, and output light is carried out optical shaping;
Described camera bellows is for covering the camera bellows of whole light path;
Described fluorescence signal receiving system comprises successively dichroic beam splitter B, optical filter, achromat B, slit B and the photomultiplier of mechanical connection; Dichroic beam splitter B is arranged between catoptron and the dichroic beam splitter A, photomultiplier and the electrical connection of Computer display control system;
Described fluorescence signal receiving system becomes confocal arrangement with the optical excitation system, and namely the slit B in the fluorescence signal receiving system lays respectively on the focus of achromat B corresponding to the place ahead and microcobjective with sample stage;
Described photoacoustic signal receiving system comprises focus supersonic detector and the amplifier that connects successively; The focus supersonic detector is connected with sample stage, the electrical connection of Amplifier And Computer display control program;
The optical focus of the microcobjective in the acoustic focus of described focus supersonic detector and the optical excitation system is confocal;
Described excitation source is preferably the short pulse excitation light source, and the effect of excitation source is exciting light acoustical signal and fluorescence signal; The excitation wavelength of described excitation source is preferably 400～2500nm, and the pulsewidth scope is preferably 1～50ns, and repetition frequency is preferably 1Hz～15KHz;
Preferred, the excitation wavelength of described excitation source is 532nm, and the pulsewidth scope is 10ns, and repetition frequency is 15Hz;
The effect of described focus supersonic detector is to receive photoacoustic signal; The effect of described photomultiplier is to receive fluorescence signal;
Described Computer display control system disposes binary channels parallel acquisition card and gathers control software, and fluorescence signal and photoacoustic signal are by the card collection of binary channels parallel acquisition and store the computing machine of controlling software with gathering into;
Use above-mentioned optoacoustic-fluorescence flow cytometer to detect the detection method of cell or particle number, may further comprise the steps:
(1) the sample fixed placement is central at sample stage, with microcobjective be placed on sample directly over, the focus supersonic detector place sample under, photomultiplier places the fluorescence light-emitting window place of microcobjective oblique upper, the photomultiplier transit tube outside is sealed with camera bellows;
(2) excitation source sends pulse laser, pass through successively diaphragm, cylindrical lens, slit A, achromat A, catoptron and microcobjective after, form a focal line hot spot and be across on the detected sample, and vertical with the flow direction of sample; The size of focal line hot spot is preferably 5 μ m * 60 μ m;
(3) when sample flow warp spot area, produce simultaneously optoacoustic and fluorescence signal owing to exciting of laser;
(4) fluorescence signal that produces through microcobjective, dichroic beam splitter B, optical filter, achromat B, and slit B after received by photomultiplier;
(5) photoacoustic signal that produces is received by the focus supersonic detector of sample stage below simultaneously;
(6) photoacoustic signal and fluorescence signal be respectively simultaneously by binary channels parallel acquisition card collection signal intensity and time waveform, after be stored in the computing machine;
(7) after to be collected the finishing, by the signal analysis and processing of computing machine, obtain respectively optoacoustic flow cytometer detection and the cell that passes through the linear light spot of fluorescence flow cytometer detection or the counting of particle in the sample;
The receive mode of the fluorescence signal described in the step (4) adopts reverse confocal receiving mode; Described reverse confocal receiving mode refers to that photomultiplier receives is fluorescence on the excitation-emission light path, and photomultiplier becomes confocal conjugate relation with detected material, namely the slit before the photomultiplier and sample be placed on respectively receive and excitation light path on optical focus on;
The receive mode of the photoacoustic signal described in the step (5) adopts the confocal receiving mode of subtend; The confocal receiving mode of described subtend refers to microcobjective in the optical excitation system and the focus supersonic detector in the photoacoustic signal receiving system in pairs to placement, and the optical focus of microcobjective overlaps with the acoustic focus of focus supersonic detector; Sample is adjusted on the confocal point by sample stage and is realized confocally exciting and receiving;
Described optoacoustic-fluorescence flow cytometer can be applicable to quantitative test and the monitoring of cell or particle.
Principle of work of the present invention: pulse laser can absorbate bulk absorption luminous energy and is produced photoacoustic signal or fluorescence signal, perhaps produces simultaneously optoacoustic and fluorescence signal.By single excitation source, produce simultaneously two kinds of signal modes, and realize simultaneously flow cytometer detection, utilize the complementarity of optoacoustic and fluorescence signal to realize high detection efficiency.The present invention's employing excites and receives photon excited, and two passages were gathered simultaneously about photoacoustic signal and fluorescence signal divided.Pulse laser is through becoming the focal line hot spot behind the optical shaping, and this linear light spot respectively with focus supersonic detector and photomultiplier before linear slots be in confocal position, greatly improved the detection efficiency of photoacoustic signal and fluorescence signal.
The present invention has following advantage and effect with respect to prior art:
(1) the present invention combines optoacoustic and fluorescence flow cytometer detection method, under the excitation of single excitation source, produce simultaneously optoacoustic and fluorescence signal, can obtain simultaneously optics and the parameters,acoustic of detected material, the detection of the size of realization cell (or particle), density, number etc.
(2) optoacoustic of the present invention-fluorescence flow cytometer utilizes optoacoustic and fluorescence signal that complementary information can be provided, and can utilize its photoacoustic signal to analyze such as dark matter, remedies the problem that existing fluorescence flow cytometer detection depends on fluorochrome label.
(3) the present invention's employing excites and receives photon excited, eliminates the non-confocal signal, only accepts target place signal, has improved system's detection sensitivity.
Description of drawings
Fig. 1 is the structural representation of the optoacoustic-fluorescence flow cytometer of embodiment 1, and wherein: 1 is excitation source, and 2 is diaphragm, and 3 is cylindrical lens, 4 is slit A, and 5 is achromat A, and 6 is catoptron, 7 is dichroic beam splitter B, and 8 is optical filter, and 9 is achromat B, 10 is slit B, and 11 is photomultiplier, and 12 is dichroic beam splitter A, 13 is charge coupled cell, and 14 is microcobjective, and 15 is sample stage, 16 is the focus supersonic detector, and 17 is amplifier, and 18 is the Computer display control system.
Fig. 2 is the focal line hot spot figure of embodiment 1.
Fig. 3 is the photoacoustic signal figure of cell of the fluorescent dye FITC mark of embodiment 2, and wherein: (a) being photoacoustic signal figure, (b) is the partial enlarged drawing of photoacoustic signal figure.
Fig. 4 is the fluorescence signal figure of cell of the fluorescent dye FITC mark of embodiment 2, and wherein: (a) being fluorescence signal figure, (b) is the partial enlarged drawing of fluorescence signal figure.
Fig. 5 is the erythrocytic photoacoustic signal figure of embodiment 3.
Fig. 6 is the erythrocytic fluorescence signal figure of embodiment 3.
Embodiment
The present invention is described in further detail below in conjunction with embodiment and accompanying drawing, but embodiments of the present invention are not limited to this.
A kind of optoacoustic-fluorescence flow cytometer as shown in Figure 1, comprises optical excitation system, fluorescence signal receiving system, photoacoustic signal receiving system, sample stage and Computer display control system; The optical excitation system is connected with fluorescence signal receiving system, sample stage, Computer display control system respectively, the photoacoustic signal receiving system is connected with sample stage, Computer display control system respectively, and the fluorescence signal receiving system is connected with the Computer display control system; The Computer display control system is electrically connected with optical excitation system, fluorescence signal receiving system, photoacoustic signal receiving system respectively;
Described optical excitation system comprises excitation source 1, diaphragm 2, cylindrical lens 3, slit A4, achromat A5, catoptron 6, dichroic beam splitter A12 and microcobjective 14; Excitation source 1 and 18 electrical connections of Computer display control system; Diaphragm 2, cylindrical lens 3, slit A4, achromat A5, catoptron 6, dichroic beam splitter A12 and microcobjective 14 be mechanical connection successively; Microcobjective 14 and sample stage 15 mechanical connections;
Described fluorescence signal receiving system comprises dichroic beam splitter B7, optical filter 8, achromat B9, slit B10 and the photomultiplier 11 that connects successively; Dichroic beam splitter B7 is arranged between catoptron 6 and the dichroic beam splitter A12, photomultiplier 11 and Computer display control system 18 mechanical connections;
Described photoacoustic signal receiving system comprises successively focus supersonic detector 16 and the amplifier (ZFL-500) 17 of mechanical connection; Focus supersonic detector 16 and sample stage 15 mechanical connections, amplifier 17 and Computer display control system 18 mechanical connections;
Described excitation source 1, diaphragm 2, cylindrical lens 3, slit A4, achromat A5, optical filter 8, achromat B9, slit B10 and photomultiplier 11 are fixed in the camera bellows with strict coaxial structure; Diaphragm 2, cylindrical lens 3, slit A4, achromat A5, achromat B9 and slit B10 can move in the camera bellows track, and output light is carried out optical shaping;
Described excitation source is the short pulse excitation light source;
Described Computer display set-up of control system has binary channels parallel acquisition card and gathers control software; Fluorescence signal and photoacoustic signal be respectively simultaneously by binary channels parallel acquisition card collection signal intensity and time waveform, after be stored in the computing machine;
The laser that excitation source 1 sends forms line spot through focusing on behind the cylindrical lens 3, pass through again slit A4, achromat A5, reflect, then focus on through dichroic beam splitter A12 and microcobjective 14 by catoptron 6, shine on the sample, the size of focal beam spot is 5 μ m * 60 μ m, as shown in Figure 2; The linear light spot is across on the detected sample, and vertical with the flow direction of sample, thereby the linear light spot can cover the xsect of whole sample, so can excited sample through all cells (particle) of this hot spot; The photoacoustic signal that cell (particle) produces is focused ultrasonic detector 16 and receives, received by photomultiplier 11 behind fluorescence signal process dichroic beam splitter B7, optical filter 8, achromat B9 and the slit B10 that produces, two kinds of signals are stored into and carry out aftertreatment in the computing machine respectively simultaneously by binary channels parallel acquisition card collection signal intensity and time waveform.
The optoacoustic of utilization embodiment 1-fluorescence flow cytometer detects the detection method of number of cells, may further comprise the steps:
(1) the rhodamine sample is placed sample stage central authorities, with microcobjective be placed on sample stage directly over, the focus supersonic detector place sample stage under, photomultiplier places the fluorescence light-emitting window place of microcobjective oblique upper, the photomultiplier transit tube outside is sealed with camera bellows;
(2) start optoacoustic-fluorescence flow cytometer, adopt titanium jewel pulsed laser as excitation source, the Output of laser wavelength is 532nm, and pulsewidth is 10ns, and repetition frequency is 15Hz; Pulse laser with above-mentioned laser instrument produces forms the linear light spot through focusing on behind the cylindrical lens, passes through behind slit A, the achromat A by mirror reflects again, then is focused to a focal beam spot through dichroic beam splitter A and microcobjective; The size of focal beam spot is 5 μ m * 60 μ m;
(3) mobile example platform, when rhodamine sample flow warp spot area, make rhodamine sample blur-free imaging on display screen, and the focal beam spot that excitation light path is formed runs through corresponding rhodamine example cross section, wherein major axis crosses the diameter of rhodamine sample, and vertical with the flow direction of FITC labeled cell in the rhodamine sample, when the cell of FITC mark in the rhodamine sample passes through focal beam spot, the rhodamine sample will be inspired corresponding photoacoustic signal and fluorescence signal;
(4) received by photomultiplier behind fluorescence signal process microcobjective, dichroic beam splitter B, optical filter, achromat B and the slit B that produces;
(5) received by the focus supersonic detector behind the photoacoustic signal process ultrasonic coupling liquid that produces;
(6) photoacoustic signal and fluorescence signal be respectively simultaneously by binary channels parallel acquisition card collection signal intensity and time waveform, after be stored in the computing machine;
(7) after to be collected the finishing, by the signal analysis and processing program of computing machine, obtain respectively the cell count of passing through the linear light spot that optoacoustic flow cytometer detection and fluorescence flow cytometer detection count;
The photoacoustic signal figure of the cell of fluorescent dye FITC mark as shown in Figure 3, wherein: (a) being photoacoustic signal figure, (b) is the partial enlarged drawing of photoacoustic signal figure; The fluorescence signal figure of the cell of fluorescent dye FITC mark as shown in Figure 4, wherein: (a) being fluorescence signal figure, (b) is the partial enlarged drawing of fluorescence signal figure; By Fig. 3 and Fig. 4 as can be known this system can realize that optoacoustic flow cytometer detection and fluorescence flow cytometer detection count the cell count by the linear light spot; Partial enlarged drawing can be found out the number by the cell of linear light spot clearly.
The optoacoustic of utilization embodiment 1-fluorescence flow cytometer detects the detection method of number of cells, may further comprise the steps:
(1) mouse ear blood vessel is placed sample stage central authorities; With microcobjective be placed on sample stage directly over, the focus supersonic detector place sample stage under, photomultiplier places the fluorescence light-emitting window place of microcobjective oblique upper, the photomultiplier transit tube outside is sealed with camera bellows;
(2) start optoacoustic-fluorescence flow cytometer, adopt titanium jewel pulsed laser as excitation source, the Output of laser wavelength is 532nm, and pulsewidth is 10ns, and repetition frequency is 15Hz; Pulse laser with above-mentioned laser instrument produces forms line spot through focusing on behind the cylindrical lens, passes through behind slit A, the achromat A by mirror reflects again, then is focused to a focal beam spot through dichroic beam splitter A and object lens; The size of focal beam spot is 5 μ m * 60 μ m;
(3) mobile example platform, make mouse ear blood vessel blur-free imaging on display screen, and the focal beam spot that excitation light path is formed is across corresponding blood vessel, wherein major axis is across the diameter of blood vessel, and vertical with erythrocytic flow direction in the blood vessel, when red blood cell passes through the focal line hot spot, will be inspired corresponding photoacoustic signal and fluorescence signal;
(4) received by photomultiplier behind fluorescence signal process microcobjective, dichroic beam splitter B, optical filter, achromat B and the slit B that produces;
(5) received by the focus supersonic detector behind the photoacoustic signal process ultrasonic coupling liquid that produces;
(6) photoacoustic signal and fluorescence signal be respectively simultaneously by binary channels parallel acquisition card collection signal intensity and time waveform, after be stored in the computing machine;
(7) after to be collected the finishing, by the signal analysis and processing program of computing machine, obtain respectively optoacoustic flow cytometer detection and fluorescence flow cytometer detection and count cell count by the linear light spot;
The erythrocytic photoacoustic signal figure of mouse ear blood vessel as shown in Figure 5, the erythrocytic fluorescence signal figure of mouse ear blood vessel is as shown in Figure 6.By Fig. 5 and Fig. 6 as can be known this system can realize that optoacoustic flow cytometer detection and fluorescence flow cytometer detection count the cell count by the linear light spot; Also embodied simultaneously the complementarity of optoacoustic flow cytometer detection and fluorescence flow cytometer detection.
Above-described embodiment is the better embodiment of the present invention; but embodiments of the present invention are not restricted to the described embodiments; other any do not deviate from change, the modification done under Spirit Essence of the present invention and the principle, substitutes, combination, simplify; all should be the substitute mode of equivalence, be included within protection scope of the present invention.
Claims (10)
1. optoacoustic-fluorescence flow cytometer is characterized in that comprising optical excitation system, fluorescence signal receiving system, photoacoustic signal receiving system, sample stage and Computer display control system; The optical excitation system is connected with fluorescence signal receiving system, sample stage, Computer display control system respectively, the photoacoustic signal receiving system is connected with sample stage, Computer display control system respectively, and the fluorescence signal receiving system is connected with the Computer display control system; The Computer display control system is electrically connected with optical excitation system, fluorescence signal receiving system, photoacoustic signal receiving system respectively;
Described optical excitation system comprises excitation source, diaphragm, cylindrical lens, slit A, achromat A, catoptron, dichroic beam splitter A and microcobjective; Excitation source and the electrical connection of Computer display control system; Diaphragm, cylindrical lens, slit A, achromat A, catoptron, dichroic beam splitter A and microcobjective be mechanical connection successively; Microcobjective and described sample stage mechanical connection;
Described fluorescence signal receiving system comprises successively dichroic beam splitter B, optical filter, achromat B, slit B and the photomultiplier of mechanical connection; Dichroic beam splitter B is arranged between catoptron and the dichroic beam splitter A, photomultiplier and the electrical connection of Computer display control system;
Described photoacoustic signal receiving system comprises focus supersonic detector and the amplifier that connects successively; The focus supersonic detector is connected with sample stage, the electrical connection of Amplifier And Computer display control program.
2. optoacoustic according to claim 1-fluorescence flow cytometer, it is characterized in that: described optical excitation system device all is fixed in the camera bellows, and diaphragm, cylindrical lens, slit A, achromat A can move in the camera bellows track; Described camera bellows is for covering the camera bellows of whole light path.
3. optoacoustic according to claim 1-fluorescence flow cytometer, it is characterized in that: described fluorescence signal receiving system becomes confocal arrangement with the optical excitation system.
4. optoacoustic according to claim 3-fluorescence flow cytometer, it is characterized in that: slit B and sample stage in the described fluorescence signal receiving system lay respectively on the focus of achromat B and microcobjective.
5. optoacoustic according to claim 1-fluorescence flow cytometer, it is characterized in that: the optical focus of the microcobjective in the acoustic focus of described focus supersonic detector and the optical excitation system is confocal.
6. optoacoustic according to claim 1-fluorescence flow cytometer, it is characterized in that: described excitation source is the short pulse excitation light source, the excitation wavelength of described short pulse excitation light source is 400～2500nm, and the pulsewidth scope is 1～50ns, and repetition frequency is 1Hz～15KHz.
7. optoacoustic according to claim 6-fluorescence flow cytometer, it is characterized in that: the excitation wavelength of described excitation source is 532nm, and the pulsewidth scope is 10ns, and repetition frequency is 15Hz.
8. optoacoustic according to claim 1-fluorescence flow cytometer is characterized in that: described Computer display control system disposes binary channels parallel acquisition card and gathers control software.
9. use the detection method of each described optoacoustic-fluorescence flow cytometer of claim 1～8, may further comprise the steps:
(1) the sample fixed placement is central at sample stage, with microcobjective be placed on sample directly over, the focus supersonic detector place sample under, photomultiplier places the fluorescence light-emitting window place of microcobjective oblique upper, the photomultiplier transit tube outside is sealed with camera bellows;
(2) excitation source sends pulse laser, pass through successively diaphragm, cylindrical lens, slit A, achromat A, catoptron and microcobjective after, form a focal line hot spot and be across on the detected sample, and vertical with the flow direction of sample; The size of focal line hot spot is 5 μ m * 60 μ m;
(3) when sample flow warp spot area, produce simultaneously optoacoustic and fluorescence signal owing to exciting of laser;
(4) fluorescence signal that produces through microcobjective, dichroic beam splitter B, optical filter, achromat B, and slit B after received by photomultiplier;
(5) photoacoustic signal that produces is received by the focus supersonic detector of sample stage below simultaneously;
(6) photoacoustic signal and fluorescence signal be respectively simultaneously by binary channels parallel acquisition card collection signal intensity and time waveform, after be stored in the computing machine;
(7) after to be collected the finishing, by the signal analysis and processing of computing machine, obtain respectively optoacoustic flow cytometer detection and the cell that passes through the linear light spot of fluorescence flow cytometer detection or the counting of particle in the sample;
The receive mode of the fluorescence signal described in the step (4) adopts reverse confocal receiving mode; The receive mode of the photoacoustic signal described in the step (5) adopts the confocal receiving mode of subtend.
10. the application of each described optoacoustic-fluorescence flow cytometer of claim 1～8, it is characterized in that: described optoacoustic-fluorescence flow cytometry is in quantitative test and the monitoring of cell or particle.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210574122.2A CN103033464B (en) | 2012-12-26 | 2012-12-26 | Photoacoustic-fluorescence flow cytometer |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210574122.2A CN103033464B (en) | 2012-12-26 | 2012-12-26 | Photoacoustic-fluorescence flow cytometer |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN103033464A true CN103033464A (en) | 2013-04-10 |
| CN103033464B CN103033464B (en) | 2014-12-24 |
Family
ID=48020584
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201210574122.2A Active CN103033464B (en) | 2012-12-26 | 2012-12-26 | Photoacoustic-fluorescence flow cytometer |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103033464B (en) |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103983578A (en) * | 2014-05-23 | 2014-08-13 | 华南师范大学 | Method and device for simultaneously microimaging scattered opto-acoustic-confocal fluorescence bimodule |
| CN104459975A (en) * | 2014-12-29 | 2015-03-25 | 中国科学院长春光学精密机械与物理研究所 | Microimaging optical system of imaging flow cytometry |
| CN105352931A (en) * | 2015-09-28 | 2016-02-24 | 周辉 | Multifunctional device and method for detecting tumor cells or other pathologic cells |
| CN104280327B (en) * | 2014-06-20 | 2017-01-04 | 博奥生物集团有限公司 | A kind of streaming phosphor collection optical system |
| CN108519329A (en) * | 2018-03-26 | 2018-09-11 | 华中科技大学 | A kind of line co-focusing imaging device of multi-channel scanning and detection |
| CN112345502A (en) * | 2020-10-27 | 2021-02-09 | 天津市英贝特航天科技有限公司 | Detector for eliminating fluorescence noise in fluorescence immunoassay analyzer |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101278829A (en) * | 2008-05-26 | 2008-10-08 | 上海理工大学 | Portable in vivo flow cytometry |
| US20090156932A1 (en) * | 2007-12-13 | 2009-06-18 | Board Of Trustees Of The University Of Arkansas | Device and method for in vivo flow cytometry using the detection of photoacoustic waves |
| CN102087198A (en) * | 2010-11-18 | 2011-06-08 | 苏州生物医学工程技术研究所 | Flow cytometry |
-
2012
- 2012-12-26 CN CN201210574122.2A patent/CN103033464B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20090156932A1 (en) * | 2007-12-13 | 2009-06-18 | Board Of Trustees Of The University Of Arkansas | Device and method for in vivo flow cytometry using the detection of photoacoustic waves |
| CN101278829A (en) * | 2008-05-26 | 2008-10-08 | 上海理工大学 | Portable in vivo flow cytometry |
| CN102087198A (en) * | 2010-11-18 | 2011-06-08 | 苏州生物医学工程技术研究所 | Flow cytometry |
Non-Patent Citations (1)
| Title |
|---|
| VLADIMIR P. ZHAROV ET AL.: "In vivo photoacoustic flow cytometry for monitoring of circulating single cancer cells and contrast agents", 《"OPTICS LETTERS》 * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103983578A (en) * | 2014-05-23 | 2014-08-13 | 华南师范大学 | Method and device for simultaneously microimaging scattered opto-acoustic-confocal fluorescence bimodule |
| CN104280327B (en) * | 2014-06-20 | 2017-01-04 | 博奥生物集团有限公司 | A kind of streaming phosphor collection optical system |
| CN104459975A (en) * | 2014-12-29 | 2015-03-25 | 中国科学院长春光学精密机械与物理研究所 | Microimaging optical system of imaging flow cytometry |
| CN105352931A (en) * | 2015-09-28 | 2016-02-24 | 周辉 | Multifunctional device and method for detecting tumor cells or other pathologic cells |
| CN108519329A (en) * | 2018-03-26 | 2018-09-11 | 华中科技大学 | A kind of line co-focusing imaging device of multi-channel scanning and detection |
| CN112345502A (en) * | 2020-10-27 | 2021-02-09 | 天津市英贝特航天科技有限公司 | Detector for eliminating fluorescence noise in fluorescence immunoassay analyzer |
Also Published As
| Publication number | Publication date |
|---|---|
| CN103033464B (en) | 2014-12-24 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US11698334B2 (en) | Flow cytometer with optical equalization | |
| KR102527830B1 (en) | Cell sorting using a high throughput fluorescence flow cytometer | |
| CN108387505B (en) | Multifunctional optical tweezers system and method based on micro-fluidic chip | |
| JP6215454B2 (en) | Image flow cytometer, system and method | |
| CN204731160U (en) | A kind of autofluorescence life-span imaging and fluorescence spectrum combine the device being used for early diagnosis of cancer | |
| CN104880445B (en) | A kind of autofluorescence life-span imaging and fluorescence spectrum combine the device for early diagnosis of cancer | |
| CN102621115B (en) | Confocal simultaneous opto-acoustic imaging and fluorescence imaging method and device | |
| CN105300943B (en) | A kind of microscope integrated optical circuit system for drop fluorescence detection | |
| WO1998059233A1 (en) | Interlaced lasers for multiple fluorescence measurement | |
| CN103487359B (en) | A kind of cell of laser excitation and particle shape and apparatus for measuring distribution | |
| CN103033464B (en) | Photoacoustic-fluorescence flow cytometer | |
| CN101278829A (en) | Portable in vivo flow cytometry | |
| CN203587475U (en) | Cell and particle morphology optical detection device | |
| CN102998293A (en) | Multichannel quantitative detection device and detection method of two-photon fluorescence optical tweezers | |
| TW201105950A (en) | System and method for the measurement of multiple emissions from multiple parallel flow channels in a flow cytometry system | |
| EP2843410B1 (en) | Sample analyzing method and sample analyzer | |
| CN104458545A (en) | Speed detection device for imaging flow cytometer | |
| JP3815838B2 (en) | Particle measuring device | |
| CN201852788U (en) | Portable fluorescence detector | |
| US11959849B2 (en) | Flow cytometers including light collection enhancers, and methods of using the same | |
| CN102095712A (en) | Photoelectric sensor for detecting blue-green algae | |
| JP5542067B2 (en) | Molecular diagnostic system based on evanescent irradiation and fluorescence | |
| CN110082330A (en) | Combined system is imaged in a kind of laser optical tweezer Raman spectrum and multi-photon | |
| CN201984016U (en) | Photoelectric sensor for detecting blue-green algae | |
| JP2022524708A (en) | Photodetection system and how to use it |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant |