CN105588827B - Living single cell Raman analysis platform digital control system and method - Google Patents
Living single cell Raman analysis platform digital control system and method Download PDFInfo
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- CN105588827B CN105588827B CN201410579147.0A CN201410579147A CN105588827B CN 105588827 B CN105588827 B CN 105588827B CN 201410579147 A CN201410579147 A CN 201410579147A CN 105588827 B CN105588827 B CN 105588827B
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Abstract
The present invention relates to unicellular sorting field, specifically a kind of Living single cell Raman analysis platform digital control system and method.System includes instrument controlling module, cell image recognition analysis module, Raman analysis processing module and unicellular screening control module；Method includes：Input sample, data acquisition, edge detection and coordinate identification, instrument controlling module are moved to the position where cell according to cell coordinate control XYZ axis；Single cell Raman spectrum and Raman spectrum data library are compared, the unicellular information is obtained；When the selection bifurcation of the unicellular arrival micro-fluidic chip for the condition that meets, unicellular push-in is screened into cell outlet using instrument controlling module control solenoid valve, otherwise unicellular inflow waste liquid pool.The application of the present invention will form systematization, complete report control platform, comprehensive support and the control section for improving high-throughput Living single cell light pincers screening system in situ, and then the research for the single cell analysis that raises speed.
Description
Technical field
The present invention relates to unicellular sorting field, specifically a kind of Living single cell Raman analysis platform digital control
System and method.
Background technology
Single cell analysis early diagnoses major disease, treats, drug screening and the research of cell physiological, pathologic process have
Significance has become one of the hot spot of research at present.Since cell is minimum, diameter is generally 5-500 μm, volume in fL-nL,
Constituent content few (fmol-zmol), type are various so that manipulate and analysis difficulty is bigger.Therefore, to carry out individual cell level
Research, the precondition that cell sorting necessitates.
Single celled identification and sorting, basis are correctly to acquire the index of characterization cellular informatics.Existing conventional flow
In formula cell instrument, the identification of cell and particle information mainly uses common optical signal, information acquisition rate of the unit interval to target
It is relatively low, therefore under the premise of meeting the recognition efficiency of engineer application, lack enough information, can only to the form of cell/particle,
The limited index such as index of refraction, reflectivity or fluorescence intensity distinguishes, and is difficult to differentiate between the cell type and physiology of active somatic cell
Characteristic, it is even more impossible to provide the gene expression of cellular level and functional information.
Living single cell Raman sorter based on Raman, may be implemented on individual cell level to having specific compound
The cell individual of collection of illustrative plates or group counted, function distinguishing and physical separation, is realized in terms of apparent type and genotype two
Investigate simultaneously and the function of verification individual cells (and its group), in scientific research chain and industry chain (supply chain) to these cells
Group credit analysis is transformed and is utilized and establishes basic basis.The machine is by laser, Raman spectrometer, epifluorescence microscope, micro-
Flow control cell sorting system forms, and is the instrument of cell sorting of the First based on cell Raman finger-print in the world.At present
The similar research equipment not being commercialized both at home and abroad.Sorter can realize unicellular Raman collection of illustrative plates acquisition, cell category at present
And growth conditions differentiate, Raman-epi-fluorescence can not cultivate functional microorganism identification, Raman light pincers Manipulation of single cells, Raman light
Clamp the functions such as unicellular sorting, the Raman signal for being based on Surface enhanced Raman scattering (SERS) technology.
Invention content
The problem of for the time for sorting unicellular appearance at present and the precision upper deviation, inefficiency, while solving existing
Cell sorting equipment it is relatively low to the information acquisition rate of target the problems such as, the technical problem to be solved in the present invention is to provide one kind can
Automation, the intelligent Living single cell Raman analysis platform number to Living single cell progress image Raman collection analysis and sorting
Word control system and method are particularly suitable for application to carry out cells in situ sorting to Living single cell.
Present invention technical solution used for the above purpose is：A kind of Living single cell Raman analysis platform digital
Control system, including：
Instrument controlling module, the parameter information for acquiring microscope, EMCCD, three-dimensional regulation platform, and will be collected
Parameter information is sent to cell image recognition analysis module, receives the cell coordinate information of cell image recognition analysis module, root
The XYZ axis amount of movements of three-dimensional regulation platform are generated according to the cell coordinate information；
Cell image recognition analysis module, for according to the collected parameter of instrument controlling module do cell edge detection,
Cell recognition and coordinate identification obtain cell coordinate, and cell coordinate are sent to instrument controlling module and Raman analysis processing
Module；
Raman analysis processing module, for scanning cell according to cell coordinate and obtaining its Raman spectrum, by Raman spectrum
The data obtained using svm classifier algorithm are compared with the data in known Raman spectrum data library, obtain unicellular kind
Class and condition of culture information send single cell species and condition of culture information to unicellular screening control module；
Unicellular screening control module, the single cell species provided according to Raman analysis processing module and condition of culture information
Unicellular screening is carried out, it is unicellular to obtain required type.
The data that Raman spectrum is obtained using svm classifier algorithm and the data in known Raman spectrum data library
In, record single celled 4 category information：A, project information, including serial number, testing time；B, sample preparation information, including cell
Name, temperature；C, instrument parameter, including laser wavelength, grating, the time of integration；D, cellular informatics, including position, image, drawing
Graceful spectrogram.
A kind of Living single cell Raman analysis platform digital control method, includes the following steps：
Input sample：It inputs different types of unicellular；
Instrument controlling module acquires microscope and the parameter of EMCCD, three-dimensional regulation platform, and collected parameter is sent
Give cell image recognition analysis module；
Cell edge detection, cell recognition and coordinate identification are done according to the collected parameter of instrument controlling module, is obtained thin
Born of the same parents' coordinate, and cell coordinate is sent to instrument controlling module and Raman analysis processing module；
Instrument controlling module is moved to the position where cell according to cell coordinate control XYZ axis；Raman analysis handles mould
Cell coordinate is supplied to Raman spectroscopy scans light to clamp by block, is scanned all cells successively and is obtained its Raman spectrum；
Single cell Raman spectrum and Raman spectrum data library are compared, the unicellular information is obtained；
When the selection bifurcation of the unicellular arrival micro-fluidic chip for the condition that meets, controlled using instrument controlling module
Unicellular push-in is screened cell outlet by solenoid valve, otherwise unicellular inflow waste liquid pool.
The cell edges detection uses Sobel/Prewitt image processing algorithms.
The cell recognition and coordinate identification are detected cell edges using image procossing mode and position acquisition.
Cell coordinate is supplied to Raman spectroscopy scans light to clamp by the Raman analysis processing module, scans all cells successively
And its Raman spectrum is obtained, specifically include following steps：
Three-dimensional regulation platform moves, and three-dimensional regulation platform is adjusted to cell position according to cell coordinate；
It controls laser to open, and controls EMCCD and carry out spectrum data gathering and preservation；
Carry out next Cellular spectroscopic information collection.
The present invention has the following advantages and beneficial effects：
1. design, exploitation and the optimization of the present invention, carry for the Single cell analysis and sorting of automation, intelligence and high throughput
Technical guarantee is supplied.
2. the application of the present invention will be formed systematization, complete (including unicellular image and Raman signal extraction etc.)
Report control platform, comprehensive support and the control section for improving high-throughput Living single cell light pincers screening system in situ, and then it is single to raise speed
The research of cell analysis.
Description of the drawings
Fig. 1 is the integrated stand composition of the present invention.
Specific implementation mode
The present invention is described in further detail with reference to the accompanying drawings and embodiments.
As shown in Figure 1, present system includes：
Instrument controlling module, the parameter information for acquiring microscope, EMCCD, three-dimensional regulation platform, and will be collected
Parameter information is sent to cell image recognition analysis module, receives the cell coordinate information of cell image recognition analysis module, root
The XYZ axis amount of movements of three-dimensional regulation platform are generated according to the cell coordinate information；
Cell image recognition analysis module, for according to the collected parameter of instrument controlling module do cell edge detection,
Cell recognition and coordinate identification obtain cell coordinate, and cell coordinate are sent to instrument controlling module and Raman analysis processing
Module；
Raman analysis processing module, for scanning cell according to cell coordinate and obtaining its Raman spectrum, by Raman spectrum
The data obtained using svm classifier algorithm are compared with the data in known Raman spectrum data library, obtain unicellular kind
Class and condition of culture information send single cell species and condition of culture information to unicellular screening control module；
Unicellular screening control module, the single cell species provided according to Raman analysis processing module and condition of culture information
Unicellular screening is carried out, it is unicellular to obtain required type.
The method of the present invention includes the following steps：
Input sample：It inputs different types of unicellular；
Instrument controlling module acquires microscope and the parameter of EMCCD, three-dimensional regulation platform, and collected parameter is sent
Give cell image recognition analysis module；
Cell edge detection, cell recognition and coordinate identification are done according to the collected parameter of instrument controlling module, is obtained thin
Born of the same parents' coordinate, and cell coordinate is sent to instrument controlling module and Raman analysis processing module；
Instrument controlling module is moved to the position where cell according to cell coordinate control XYZ axis；Raman analysis handles mould
Cell coordinate is supplied to Raman spectroscopy scans light to clamp by block, is scanned all cells successively and is obtained its Raman spectrum；
Single cell Raman spectrum and Raman spectrum data library are compared, the unicellular information is obtained；
When the selection bifurcation of the unicellular arrival micro-fluidic chip for the condition that meets, controlled using instrument controlling module
Unicellular push-in is screened cell outlet by solenoid valve, otherwise unicellular inflow waste liquid pool.
The cell edges detection uses Sobel/Prewitt image processing algorithms.
The cell recognition and coordinate identification are detected cell edges using image procossing mode and position acquisition.Figure
As processing mode includes greyscale transformation, smooth, edge extracting, to realize single celled positioning.
Cell coordinate is supplied to Raman spectroscopy scans light to clamp by the Raman analysis processing module, scans all cells successively
And its Raman spectrum is obtained, specifically include following steps：
Three-dimensional regulation platform moves, and three-dimensional regulation platform is adjusted to cell position according to cell coordinate；
It controls laser to open, and controls EMCCD and carry out spectrum data gathering and preservation；
Carry out next Cellular spectroscopic information collection.
Claims (6)
1. a kind of Living single cell Raman analysis platform digital control system, which is characterized in that including：
Instrument controlling module, the parameter information for acquiring microscope, EMCCD, three-dimensional regulation platform, and by collected parameter
Information is sent to cell image recognition analysis module, the cell coordinate information of cell image recognition analysis module is received, according to institute
State the XYZ axis amount of movements that cell coordinate information generates three-dimensional regulation platform；
Cell image recognition analysis module, for doing cell edge detection, cell according to the collected parameter of instrument controlling module
Identification and coordinate identification, obtain cell coordinate, and cell coordinate is sent to instrument controlling module and Raman analysis processing module；
Raman analysis processing module utilizes Raman spectrum for scanning cell according to cell coordinate and obtaining its Raman spectrum
The data that svm classifier algorithm obtains are compared with the data in known Raman spectrum data library, obtain single cell species and
Condition of culture information sends single cell species and condition of culture information to unicellular screening control module；
Unicellular screening control module, the single cell species provided according to Raman analysis processing module and condition of culture information carry out
Unicellular screening, it is unicellular to obtain required type.
2. Living single cell Raman analysis platform digital control system according to claim 1, which is characterized in that the drawing
The data that graceful spectrum is obtained using svm classifier algorithm and the data in the known Raman spectrum data library, record slender
4 category informations of born of the same parents：A, project information, including serial number, testing time；B, sample preparation information, including cell name, temperature；C、
Instrument parameter, including laser wavelength, grating, the time of integration；D, cellular informatics, including position, image, raman spectrum.
3. a kind of Living single cell Raman analysis platform digital control method, which is characterized in that include the following steps：
Input sample：It inputs different types of unicellular；
Instrument controlling module acquires microscope and the parameter of EMCCD, three-dimensional regulation platform, and collected parameter is sent to carefully
Born of the same parents' image recognition analysis module；
Cell edge detection, cell recognition and coordinate identification are done according to the collected parameter of instrument controlling module, obtains cell seat
Mark, and cell coordinate is sent to instrument controlling module and Raman analysis processing module；
Instrument controlling module is moved to the position where cell according to cell coordinate control XYZ axis；Raman analysis processing module will
Cell coordinate is supplied to Raman spectroscopy scans light to clamp, and scans all cells successively and obtains its Raman spectrum；
Single cell Raman spectrum and Raman spectrum data library are compared, the unicellular information is obtained；
When the selection bifurcation of the unicellular arrival micro-fluidic chip for the condition that meets, electromagnetism is controlled using instrument controlling module
Unicellular push-in is screened cell outlet by valve, otherwise unicellular inflow waste liquid pool.
4. Living single cell Raman analysis platform digital control method according to claim 3, which is characterized in that described thin
Born of the same parents' edge detection uses Sobel/Prewitt image processing algorithms.
5. Living single cell Raman analysis platform digital control method according to claim 3, which is characterized in that described thin
Born of the same parents identify and coordinate identification is detected to cell edges using image procossing mode and position acquisition.
6. Living single cell Raman analysis platform digital control method according to claim 3, which is characterized in that the drawing
Cell coordinate is supplied to Raman spectroscopy scans light to clamp by graceful analysis and processing module, is scanned all cells successively and is obtained its Raman light
Spectrum, specifically includes following steps：
Three-dimensional regulation platform moves, and three-dimensional regulation platform is adjusted to cell position according to cell coordinate；
It controls laser to open, and controls EMCCD and carry out spectrum data gathering and preservation；
Carry out next Cellular spectroscopic information collection.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109001180A (en) * | 2018-08-10 | 2018-12-14 | 青岛启明生物科技有限公司 | A kind of Raman spectrum combination artificial intelligence high throughput single cell analysis identification method |
Families Citing this family (7)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN106769693A (en) * | 2016-11-14 | 2017-05-31 | 中国科学院重庆绿色智能技术研究院 | A kind of circulating tumor cell automatic checkout system based on Raman spectrum |
| CN108020320B (en) * | 2017-12-26 | 2023-08-29 | 同方威视技术股份有限公司 | Raman spectrum detection equipment and method based on image recognition |
| CN109266717B (en) * | 2018-09-27 | 2022-02-22 | 上海镭立激光科技有限公司 | Method and device for detecting bacterial drug resistance through single cell analysis |
| CN109943475A (en) * | 2019-04-12 | 2019-06-28 | 广西医科大学第一附属医院 | The micro-fluidic sorting chip of one kind and its separation system |
| CN110331125B (en) * | 2019-06-28 | 2023-04-07 | 南京航空航天大学 | Method for automatically realizing pairing of multiple groups of cells |
| CN112308892A (en) * | 2020-11-16 | 2021-02-02 | 中国海洋大学 | Shell texture analysis system and method based on LIBS technology |
| CN114317249A (en) * | 2022-03-15 | 2022-04-12 | 南方海洋科学与工程广东省实验室(广州) | Marine in-situ environment single cell high-throughput sorting device and method |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1804593A (en) * | 2006-01-18 | 2006-07-19 | 中国科学院上海光学精密机械研究所 | Method for distinguishing epithelial carcinoma property by single cell Raman spectrum |
| CN102019277A (en) * | 2010-10-29 | 2011-04-20 | 北京惟馨雨生物科技有限公司 | Sorter and sorting method for separating cells and particles |
| CN102183504A (en) * | 2011-01-25 | 2011-09-14 | 山东师范大学 | Microfluidic unicellular active oxygen automatic analyzer |
| CN103353452A (en) * | 2013-07-12 | 2013-10-16 | 北京惟馨雨生物科技有限公司 | Cell carrier chip and single cell rapid identifying or sorting method employing same |
| CN103940801A (en) * | 2014-04-17 | 2014-07-23 | 中国科学院青岛生物能源与过程研究所 | Method for rapid identification of microalgae on single cell level |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US7570356B2 (en) * | 2006-01-05 | 2009-08-04 | Chemimage Corporation | System and method for classifying cells and the pharmaceutical treatment of such cells using Raman spectroscopy |
| JP6236005B2 (en) * | 2011-08-30 | 2017-11-22 | バテル メモリアル インスティチュート | Identification of mycoplasma contamination using Raman spectroscopy in biotechnology production |
-
2014
- 2014-10-24 CN CN201410579147.0A patent/CN105588827B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1804593A (en) * | 2006-01-18 | 2006-07-19 | 中国科学院上海光学精密机械研究所 | Method for distinguishing epithelial carcinoma property by single cell Raman spectrum |
| CN102019277A (en) * | 2010-10-29 | 2011-04-20 | 北京惟馨雨生物科技有限公司 | Sorter and sorting method for separating cells and particles |
| CN102183504A (en) * | 2011-01-25 | 2011-09-14 | 山东师范大学 | Microfluidic unicellular active oxygen automatic analyzer |
| CN103353452A (en) * | 2013-07-12 | 2013-10-16 | 北京惟馨雨生物科技有限公司 | Cell carrier chip and single cell rapid identifying or sorting method employing same |
| CN103940801A (en) * | 2014-04-17 | 2014-07-23 | 中国科学院青岛生物能源与过程研究所 | Method for rapid identification of microalgae on single cell level |
Non-Patent Citations (1)
| Title |
|---|
| 苜蓿抗性变异细胞系的筛选;李波等;《草业科学》;20030430;第20卷(第4期);第5-9页 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109001180A (en) * | 2018-08-10 | 2018-12-14 | 青岛启明生物科技有限公司 | A kind of Raman spectrum combination artificial intelligence high throughput single cell analysis identification method |
| CN109001180B (en) * | 2018-08-10 | 2021-01-01 | 青岛启明生物科技有限公司 | Raman spectrum combined artificial intelligence high-throughput single cell analysis and identification method |
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Effective date of registration: 20210727 Address after: 266101 Shandong Province, Qingdao city Laoshan District Songling Road No. 189 Patentee after: Qingdao xingsai Biotechnology Co.,Ltd. Address before: 266101 Shandong Province, Qingdao city Laoshan District Songling Road No. 189 Patentee before: Qingdao Institute of Bioenergy and Process, Chinese Academy of Sciences |