CN104808327A - Microscope used for cell manipulation - Google Patents
Microscope used for cell manipulation Download PDFInfo
- Publication number
- CN104808327A CN104808327A CN201510249312.0A CN201510249312A CN104808327A CN 104808327 A CN104808327 A CN 104808327A CN 201510249312 A CN201510249312 A CN 201510249312A CN 104808327 A CN104808327 A CN 104808327A
- Authority
- CN
- China
- Prior art keywords
- laser
- microscope
- mems
- galvanometer
- mono
- 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
- 239000012472 biological sample Substances 0.000 claims abstract description 17
- MURGITYSBWUQTI-UHFFFAOYSA-N fluorescin Chemical compound OC(=O)C1=CC=CC=C1C1C2=CC=C(O)C=C2OC2=CC(O)=CC=C21 MURGITYSBWUQTI-UHFFFAOYSA-N 0.000 claims description 8
- 230000001276 controlling effect Effects 0.000 claims description 4
- 230000001105 regulatory effect Effects 0.000 claims description 4
- 230000003287 optical effect Effects 0.000 abstract 2
- 102000034287 fluorescent proteins Human genes 0.000 abstract 1
- 108091006047 fluorescent proteins Proteins 0.000 abstract 1
- 210000004027 cell Anatomy 0.000 description 19
- 238000000034 method Methods 0.000 description 10
- 238000011160 research Methods 0.000 description 4
- 230000001413 cellular effect Effects 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 230000001272 neurogenic effect Effects 0.000 description 2
- 238000005424 photoluminescence Methods 0.000 description 2
- 239000000523 sample Substances 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 230000000638 stimulation Effects 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 238000013481 data capture Methods 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000005284 excitation Effects 0.000 description 1
- 239000007850 fluorescent dye Substances 0.000 description 1
- 238000001215 fluorescent labelling Methods 0.000 description 1
- 230000004927 fusion Effects 0.000 description 1
- 239000004519 grease Substances 0.000 description 1
- 238000003384 imaging method Methods 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 239000002184 metal Substances 0.000 description 1
- 210000003061 neural cell Anatomy 0.000 description 1
- 210000002569 neuron Anatomy 0.000 description 1
- 229920001296 polysiloxane Polymers 0.000 description 1
- 210000004895 subcellular structure Anatomy 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 230000000451 tissue damage Effects 0.000 description 1
- 231100000827 tissue damage Toxicity 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
- 230000003313 weakening effect Effects 0.000 description 1
Abstract
The invention relates to a microscope used for cell manipulation. The microscope comprises a laser transmitter, which is used for transmitting laser, and an optical path system, which is connected to the laser transmitter and is used for enabling the laser to pass and exciting a biological sample to emit fluorescence or activate fluorescent protein, and further comprises an MEMS (micro-Electro-Mechanical System) single oscillating mirror, which is arranged between the laser transmitter and the optical path system and is used for controlling the laser for scanning or exciting a specified point of the biological sample in order to realize manipulation of the cells in the biological sample. In comparison with the prior art, the MEMS single oscillating mirror controls the laser for scanning or exciting the point of the biological sample, so that a laser ejection angle can be precisely adjusted, the equipment resolution is improved to a unicellular level, meanwhile, more than 90% of laser beam is utilized, and the microscope has the advantages of high resolution, high light utilization rate and low cost.
Description
Technical field
The present invention relates to a kind of cell analysis equipment, especially relate to a kind of microscope for cell operation.
Background technology
In recent years, along with the development of laser technology and microtechnic, people are also deep into Manipulation of single cells aspect to the understanding of life science, usually need to carry out microoperation to individual cells, as controlled the specificity of cell in the research of related Neurons in neurogenic disease, light genetics experiments in biological and medical research.Therefore, high-precision cell operation equipment is essential.
In cell operation experiment, the main method adopted has Writhing test, electrostimulation and light stimulus method at present.The physicochemical property of chemical stimulation method to intraor extracellular environment has considerable influence, and reach is large, is not suitable for accurate cell operation; Although the intensity of electro photoluminescence in electrical stimulation method, waveform and duration easily control, reusable, not easily cause tissue damage, cytoactive is had a certain impact, and the intensity of electro photoluminescence needed for different cell is different, adds enforcement difficulty; And the physicochemical environment of light stimulus method hardly inside and outside interference cell and its normal vital movement, and accurately can control in time, space, the manipulation of high-precision cell can be realized.
Although common fluorescent microscope is also utilize light stimulus method, its light beam irradiation area is large, is difficult to the unicellular manipulation realizing high score rate.Current state-of-the-art digital micromirror array (DMD) cellular micromanipulation equipment, DMD is utilized to segment light, resolution is at 11 microns, but DMD has certain weakening effect for excitation source, therefore high-power laser beam is required, in addition, the DMD of high pixel density manufactures difficulty and makes its resolution limitations.The most frequently used Laser Scanning Confocal Microscope utilizes laser scanning system to realize high-precision cellular micromanipulation, but it has confocal system, be mainly used in three-dimensional imaging, and the accurate test to subcellular structure and dynamic process, and serviceable life is limited, expensive, complex structure, operation easier is large, is difficult to popularize.
Summary of the invention
Object of the present invention be exactly in order to overcome above-mentioned prior art exist defect and a kind of microscope for cell operation is provided.
Object of the present invention can be achieved through the following technical solutions:
For a microscope for cell operation, comprising:
Generating laser, for Emission Lasers;
Light path system, is connected with generating laser, passes through and excite biological sample send fluorescence or activate fluorescin for laser;
Also comprise:
The mono-galvanometer of MEMS, is located between generating laser and light path system, scans for controlling laser or excites biological sample specified point, to realize handling the cell in biological sample.
Described microscope also comprises the ccd image sensor be connected with light path system, and the control module be connected with the mono-galvanometer of MEMS, and described control module is also connected with ccd image sensor, and:
Receive the fluoroscopic image formed by gathering fluorescence by ccd image sensor, and regulate the vibration of the mono-galvanometer of MEMS and deflection state to excite to select the specified point on biological sample according to this fluoroscopic image.
Described control module comprises the computing unit be connected with ccd image sensor, and the data collecting card be connected with the mono-galvanometer of MEMS, and described computing unit is connected with data collecting card, and
Calculating according to fluoroscopic image and the specified point that will scan or excite the coordinate information generating this specified point, coordinate information being converted to the drive singal for regulating the mono-galvanometer vibration of MEMS and deflection state by described data collecting card.
Described drive singal comprises the first driving electric signal and the second driving electric signal that are respectively used to drive the fast axle of the mono-galvanometer of described MEMS and slow axis.
Described first drives electric signal and second to drive electric signal to be sine voltage signal, and both frequencies are different.
Described ccd image sensor is located at the observation panel place above microscope ocular.
Described laser is microbeam laser.
Described microscope also comprises the fixed support for fixed laser transmitter and the mono-galvanometer of MEMS, and this fixed support is connected with the lamp box interface on microscope.
Described generating laser comprises red laser, blue laser and green laser, and described red laser, blue laser and green laser are all connected with light path system by the mono-galvanometer of MEMS.
Compared with prior art, the present invention has the following advantages:
1) the present invention controls laser to biological sample analyzing spot or shot point by the mono-galvanometer of MEMS, can fine adjustment laser injection angle, improve device resolution to unicellular level, achieve the utilization to laser beam more than 90% simultaneously, resolution is high, and light utilization efficiency is high, cost is low.
2) ccd image sensor and control module provide the window that operator carries out for the mono-galvanometer of MEMS controlling, and have larger degree of freedom.
3) by the fluoroscopic image that formed by CCD by the algorithm of optimal design in computing unit and self-programmed software control data capture card output signal, realize the automatic conversion of coordinate signal and electric signal, and the automatic control to galvanometer deflection, thus realize laser to unicellular precise manipulation, the a series of Automated condtrol process simplification flow process of experimental implementation, greatly reduces the difficulty of equipment precise manipulation.
4) electric signal comprises the first driving electric signal and the second driving electric signal that are respectively used to drive the fast axle of the mono-galvanometer of MEMS and slow axis, Signal separator.
5) laser is microbeam laser, further increases resolution.
Accompanying drawing explanation
Fig. 1 is structural representation of the present invention;
Fig. 2 is laser galvanometer modular construction schematic diagram;
Fig. 3 is the light path schematic diagram of a complete set of equipment;
Fig. 4 is the signal Transformation Graphs of a complete set of equipment;
Wherein: 1, generating laser, 2, the mono-galvanometer of MEMS, 3, microscope body, 4, ccd image sensor, 5, data collecting card, 6, light path system, 7, laser.
Embodiment
Below in conjunction with the drawings and specific embodiments, the present invention is described in detail.The present embodiment is implemented premised on technical solution of the present invention, give detailed embodiment and concrete operating process, but protection scope of the present invention is not limited to following embodiment.
For a microscope for cell operation, comprise as shown in Figure 1 to Figure 3:
Generating laser 1, for Emission Lasers 7, this laser 7 is microbeam laser, and microbeam laser adopts collimation lens to carry out optically focused;
Light path system 6, is connected with generating laser 1, passes through and excite biological sample send fluorescence or activate fluorescin for laser 7;
Also comprise:
The mono-galvanometer of MEMS 2 (i.e. the micro electronmechanical single galvanometer of Electromagnetic Drive), is located between generating laser 1 and light path system 6, scans or excites, to realize handling the cell in biological sample for controlling laser 7 pairs of biological sample specified points.
Microscope also comprises the ccd image sensor 4 be connected with light path system 6, and the control module to be connected with the mono-galvanometer 2 of MEMS, control module is also connected with ccd image sensor 4, and receive the fluoroscopic image formed by gathering fluorescence by ccd image sensor 4, and the vibration of the mono-galvanometer 2 of MEMS and deflection state is regulated to excite to select the specified point on biological sample according to this fluoroscopic image.
Control module comprises the computing unit be connected with ccd image sensor 4, and the data collecting card 5 to be connected with the mono-galvanometer 2 of MEMS, computing unit is connected with data collecting card 5, and calculate according to fluoroscopic image and the specified point that will scan or excite the coordinate information generating this specified point, and transfer to data collecting card 5, coordinate information is converted to for regulating the mono-galvanometer 2 of MEMS to vibrate and the drive singal of deflection state by data collecting card 5; Drive singal comprises the first driving electric signal and the second driving electric signal that are respectively used to drive the fast axle of the mono-galvanometer 2 of MEMS and slow axis, first drives electric signal and second to drive electric signal to be sine voltage signal, and both frequencies are different, carry out orientation deflection by the frequency and peak value changing sine voltage signal to the mono-galvanometer 2 of MEMS to control, producing first drives electric signal and second to drive the driving circuit of electric signal to be the adjustable constant current signal of signal, and comprises the heat-conducting silicone grease being connected with sheet metal to carry out dispelling the heat.
Ccd image sensor 4 is located at the observation panel place above microscope ocular, and microscope also comprises the fixed support for fixed laser transmitter 1 and the mono-galvanometer 2 of MEMS, and this fixed support is connected with the lamp box interface on microscope.
Microbeam laser 7 is by the mono-galvanometer 2 of MEMS and the light path system 6 after improving, sample is excited to send fluorescence or activate fluorescin, ccd image sensor 4 is utilized to carry out image acquisition, the control of the mono-galvanometer 2 of MEMS is that the signal utilizing the data collecting card 5 of software control to export realizes, the laser galvanometer assembly that the mono-galvanometer of MEMS 2 and generating laser 1 form achieves the scanning of laser 7 on biological specimen or fixed point excites, by software, the mono-galvanometer 2 of MEMS is controlled, laser pointwise can be realized, scan along the line, comprehensively.
Microbeam laser deflects light source by the mono-galvanometer 2 of MEMS and enters microscope, the control signal data collecting card of the mono-galvanometer 2 of MEMS or other USB export control signal module and export, data collecting card 5 is connected with computing machine, and user selects the region that will scan or point by computer software.
Generating laser 1 comprises red laser, blue laser and green laser, red laser, blue laser and green laser are all connected with light path system 6 by the mono-galvanometer 2 of MEMS, and its laser launched enters microscope body 3 after the mono-galvanometer 2 of MEMS deflects.
The mono-galvanometer 2 of MEMS is controlled by data collecting card 5, computing machine inputs deflection voltage signal to the mono-galvanometer 2 of MEMS by described data collecting card 5, when inputting the sinusoidal voltage of 7700Hz, the mono-galvanometer 2 of MEMS scans along quick shaft direction, when inputting the sinusoidal voltage of 700Hz, the mono-galvanometer 2 of MEMS is along slow-axis direction scanning, and when to input 7700Hz and 700Hz two kinds of sinusoidal voltages simultaneously, the mono-galvanometer of MEMS 2 is in the interscan of whole galvanometer plane.
This microscope can realize the accurate manipulation to biological cell sample, the Bioexperiment such as light science of heredity, fluorescence labeling, induced with laser Fusion of Cells can be carried out, the fields such as scientific research, teaching, medical treatment can be widely used in, all there is facilitation to the research etc. of associated neural cells and neurogenic disease in Neuscience, bio-science and medical domain, solve the problem that existing equipment complicated operation, price are high.
Claims (9)
1., for a microscope for cell operation, comprising:
Generating laser (1), for Emission Lasers (7);
Light path system (6), is connected with generating laser (1), passes through and excite biological sample send fluorescence or activate fluorescin for laser (7);
It is characterized in that, also comprise:
The mono-galvanometer of MEMS (2), be located between generating laser (1) and light path system (6), for controlling laser (7), biological sample specified point is scanned or excited, to realize handling the cell in biological sample.
2. a kind of microscope for cell operation according to claim 1, it is characterized in that, described microscope also comprises the ccd image sensor (4) be connected with light path system (6), and the control module to be connected with the mono-galvanometer of MEMS (2), described control module is also connected with ccd image sensor (4), and:
Receive the fluoroscopic image formed by gathering fluorescence by ccd image sensor (4), and regulate the vibration of the mono-galvanometer of MEMS (2) and deflection state to excite to select the specified point on biological sample according to this fluoroscopic image.
3. a kind of microscope for cell operation according to claim 2, it is characterized in that, described control module comprises the computing unit be connected with ccd image sensor (4), and the data collecting card (5) to be connected with the mono-galvanometer of MEMS (2), described computing unit is connected with data collecting card (5), and
Calculating according to fluoroscopic image and the specified point that will scan or excite the coordinate information generating this specified point, coordinate information being converted to the drive singal for regulating the mono-galvanometer of MEMS (2) vibration and deflection state by described data collecting card (5).
4. a kind of microscope for cell operation according to claim 3, is characterized in that, described drive singal comprises the first driving electric signal and the second driving electric signal that are respectively used to drive the fast axle of the mono-galvanometer of described MEMS (2) and slow axis.
5. a kind of microscope for cell operation according to claim 4, is characterized in that, described first drives electric signal and second to drive electric signal to be sine voltage signal, and both frequencies are different.
6. according to a kind of microscope for cell operation in claim 1-5 described in arbitrary, it is characterized in that, described ccd image sensor (4) is located at the observation panel place above microscope ocular.
7. according to a kind of microscope for cell operation in claim 1-5 described in arbitrary, it is characterized in that, described laser (7) is microbeam laser.
8. according to a kind of microscope for cell operation in claim 1-5 described in arbitrary, it is characterized in that, described microscope also comprises the fixed support for fixed laser transmitter (1) and the mono-galvanometer of MEMS (2), and this fixed support is connected with the lamp box interface on microscope.
9. according to a kind of microscope for cell operation in claim 1-5 described in arbitrary, it is characterized in that, described generating laser (1) comprises red laser, blue laser and green laser, and described red laser, blue laser and green laser are all connected with light path system (6) by the mono-galvanometer of MEMS (2).
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510249312.0A CN104808327B (en) | 2015-05-15 | 2015-05-15 | A kind of microscope for cell operation |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510249312.0A CN104808327B (en) | 2015-05-15 | 2015-05-15 | A kind of microscope for cell operation |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN104808327A true CN104808327A (en) | 2015-07-29 |
| CN104808327B CN104808327B (en) | 2017-10-27 |
Family
ID=53693297
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510249312.0A Active CN104808327B (en) | 2015-05-15 | 2015-05-15 | A kind of microscope for cell operation |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104808327B (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108732736A (en) * | 2017-04-24 | 2018-11-02 | 复旦大学 | It is integrated with microscopic system of the digital micromirror array to electro-optical device |
Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1664558A (en) * | 2005-03-24 | 2005-09-07 | 华中科技大学 | Minisize three-dimensional self-scanning confocal microscope |
| CN101266335A (en) * | 2007-03-12 | 2008-09-17 | 精工爱普生株式会社 | Actuator, optical scanner, and image forming apparatus |
| CN101546104A (en) * | 2008-03-28 | 2009-09-30 | 船井电机株式会社 | Projection image display apparatus |
| CN102985866A (en) * | 2010-06-23 | 2013-03-20 | 浜松光子学株式会社 | Image generation device |
| CN103616760A (en) * | 2013-11-26 | 2014-03-05 | 中国科学院苏州生物医学工程技术研究所 | Laser scanning confocal microscope imaging system |
| CN103645136A (en) * | 2013-11-22 | 2014-03-19 | 深圳先进技术研究院 | Method and device for improving multiphoton fluorescence microscope imaging resolution |
| US20140247976A1 (en) * | 2011-11-10 | 2014-09-04 | Olympus Corporation | Image Analysis Method and Image Analysis Apparatus |
-
2015
- 2015-05-15 CN CN201510249312.0A patent/CN104808327B/en active Active
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1664558A (en) * | 2005-03-24 | 2005-09-07 | 华中科技大学 | Minisize three-dimensional self-scanning confocal microscope |
| CN101266335A (en) * | 2007-03-12 | 2008-09-17 | 精工爱普生株式会社 | Actuator, optical scanner, and image forming apparatus |
| CN101546104A (en) * | 2008-03-28 | 2009-09-30 | 船井电机株式会社 | Projection image display apparatus |
| CN102985866A (en) * | 2010-06-23 | 2013-03-20 | 浜松光子学株式会社 | Image generation device |
| US20140247976A1 (en) * | 2011-11-10 | 2014-09-04 | Olympus Corporation | Image Analysis Method and Image Analysis Apparatus |
| CN103645136A (en) * | 2013-11-22 | 2014-03-19 | 深圳先进技术研究院 | Method and device for improving multiphoton fluorescence microscope imaging resolution |
| CN103616760A (en) * | 2013-11-26 | 2014-03-05 | 中国科学院苏州生物医学工程技术研究所 | Laser scanning confocal microscope imaging system |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108732736A (en) * | 2017-04-24 | 2018-11-02 | 复旦大学 | It is integrated with microscopic system of the digital micromirror array to electro-optical device |
Also Published As
| Publication number | Publication date |
|---|---|
| CN104808327B (en) | 2017-10-27 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| JP5534214B2 (en) | Flow cytometer and flow cytometry method | |
| US5795755A (en) | Method of implanting living cells by laser poration at selected sites | |
| EP1835323B1 (en) | Fluorescence microscope and observation method | |
| CN104081250A (en) | Light scanning observation device | |
| US20170160200A1 (en) | Optical analysis device | |
| CN104198458A (en) | Femtosecond laser two-photon fluorescent biological microimaging system and imaging method thereof | |
| Supekar et al. | Two-photon laser scanning microscopy with electrowetting-based prism scanning | |
| CN109470666B (en) | Near infrared laser confocal biochip scanner | |
| CN105527261B (en) | A kind of microscopical multi-modal scanning means of two-photon fluorescence | |
| Lin et al. | Microfluidic cell counter/sorter utilizing multiple particle tracing technique and optically switching approach | |
| EP1512963B1 (en) | Fluorescence lifetime distribution image measuring system and its measuring method | |
| CN203164118U (en) | A fluorescence lifetime microimaging system | |
| US20220034785A1 (en) | Cell sorting device and method | |
| CN108333151B (en) | Super-resolution microscopic imaging system and method based on femtosecond pulse shaping | |
| CN107028590B (en) | Miniaturized self-adaptive optical two-photon fluorescence imaging system and method | |
| CN109612388B (en) | Optical measurement system and method for cell perforation | |
| US9341919B2 (en) | Methods and apparatus for controling drive frequencies of acousto-optic deflectors | |
| US20230175950A1 (en) | Light Detection Systems Having A Secondary Light Scatter Detector And Methods For Using Same | |
| CN116670492A (en) | Method and microscope for recording trajectories of individual particles in a sample | |
| CN114298212B (en) | Monitoring device for cell micro-loss induction and bright field monitoring method | |
| CN104808327A (en) | Microscope used for cell manipulation | |
| CN109211855B (en) | Multi-beam multi-photon microscopic imaging device | |
| EP3397770B1 (en) | Method for the optoinjection of exogenous material into a biological cell | |
| CN106338546A (en) | High-spatial-resolution imaging mass spectrometry device | |
| CN205620304U (en) | Two -photon fluorescence microscope's multimode scanning device |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| EXSB | Decision made by sipo to initiate substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| OL01 | Statement of open license |