CN101782518A - Cell opto-acoustic microscopic imaging method and device thereof - Google Patents
Cell opto-acoustic microscopic imaging method and device thereof Download PDFInfo
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- CN101782518A CN101782518A CN 201010115730 CN201010115730A CN101782518A CN 101782518 A CN101782518 A CN 101782518A CN 201010115730 CN201010115730 CN 201010115730 CN 201010115730 A CN201010115730 A CN 201010115730A CN 101782518 A CN101782518 A CN 101782518A
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N29/00—Investigating or analysing materials by the use of ultrasonic, sonic or infrasonic waves; Visualisation of the interior of objects by transmitting ultrasonic or sonic waves through the object
- G01N29/04—Analysing solids
- G01N29/06—Visualisation of the interior, e.g. acoustic microscopy
- G01N29/0654—Imaging
- G01N29/0681—Imaging by acoustic microscopy, e.g. scanning acoustic microscopy
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N29/00—Investigating or analysing materials by the use of ultrasonic, sonic or infrasonic waves; Visualisation of the interior of objects by transmitting ultrasonic or sonic waves through the object
- G01N29/22—Details, e.g. general constructional or apparatus details
- G01N29/24—Probes
- G01N29/2418—Probes using optoacoustic interaction with the material, e.g. laser radiation, photoacoustics
Abstract
The invention relates to a cell opto-acoustic microscopic imaging method, which comprises the following steps: placing a cell on the concave surface of microscopic glass, irradiating the cell with laser light permeating the microscopic glass, producing opto-acoustic effect after the cell absorbs the light, transferring acoustic pressure variation produced by the opto-acoustic effect to an opto-acoustic sensor, and detecting the opto-acoustic variation and outputting an opto-acoustic signal by using the opto-acoustic sensor to realize opto-acoustic detection of the single cell; and performing two-dimensional scanning on the cell by using light beam scanning and a micro objective with high resolution to realize the opto-acoustic microscopic imaging of the single cell. The invention also relates to a cell opto-acoustic microscopic imaging device, which comprises a laser light scanning imaging mechanism, the opto-acoustic sensor and a signal processor. In the method and the device, light beam scanning technology without mechanical noise is combined with the micro objective with the high resolution to generate the opto-acoustic signal with high spatial resolution and then the opto-acoustic sensor is adopted for the opto-acoustic detection to perform the opto-acoustic microscopic imaging on the cell, wherein the solution is less than 1 micron.
Description
Technical field
The present invention relates to micro-imaging technique, specifically is a kind of cell opto-acoustic microscopic imaging method, the invention still further relates to the cell opto-acoustic microscopic imaging device.
Background technology
Photoacoustic imaging is to adopt the method for " optical excitation-induce photoacoustic signal-optoacoustic detection-image reconstruction " to carry out imaging.The photoacoustic imaging technology has significant advantage at aspects such as image resolution ratio, contrast and quantity of information than many traditional image technologies, and acousto-optic imaging method mainly comprises following three kinds of formation methods at present: acoustic lens imaging method, phase-control focusing imaging method and inversion imaging method.The acoustic lens imaging method is owing to be subjected to the restriction of acoustic wave diffraction effect harmony lens aberration, and resolution can only reach 1 millimeter level; The phase-control focusing imaging method is to utilize polynary linear array ultrasonic transducer to carry out image reconstruction in conjunction with the phase-control focusing algorithm, and its resolution has reached 100 microns, can carry out imaging to blood vessel; The inversion imaging method is to utilize Radon inversion algorithm and filter back-projection algorithm to carry out image reconstruction, owing to avoided the restriction of acoustic wave diffraction effect, thereby can realize the photoacoustic imaging of high-resolution, its resolution has reached 15 microns, can carry out imaging to capillary.But this resolution still can't be differentiated cell, more can't carry out imaging by pair cell.
Because the biological behaviour characteristic informations such as (as growth, apoptosis, metabolism, pathology, sudden changes) that the photoacoustic imaging technology can obtain to organize, thereby realize the biological function imaging.Yet, because at present the resolution of photoacoustic imaging technology is also lower, can only reach 15 microns resolution, structural images and function image that can't the observation of cell level.
Summary of the invention
The purpose of this invention is to provide a kind of high resolving power cell opto-acoustic microscopic imaging method that can observe cellular level, realize the high resolving power opto-acoustic microscopic imaging, resolution is less than 1 micron.
The present invention also aims to provide a kind of cell opto-acoustic microscopic imaging device.
Cell opto-acoustic microscopic imaging method of the present invention comprises:
---cell is placed on the concave surface of cover glass, laser sees through cover glass and is radiated on the cell, the cell absorbing light produces optoacoustic effect, be delivered to photoacoustic sensors by the sound pressure variations that this optoacoustic effect produced, survey and export photoacoustic signal then by photoacoustic sensors, to unicellular realization optoacoustic detection;
---by beam flying and high-resolution microcobjective, pair cell carries out two-dimensional scan, realizes single celled opto-acoustic microscopic imaging.
Cell opto-acoustic microscopic imaging device of the present invention comprises laser scanning imaging mechanism, photoacoustic sensors and signal processor three parts.Wherein, laser scanning imaging mechanism is made of laser instrument, scanning galvanometer, microcobjective; Signal processor is electrically connected successively by chopper, lock-in amplifier, capture card and computing machine and constitutes.
As shown in Figure 1, the laser that laser instrument 1 sends, modulate by chopper 2, enter scanning galvanometer 3 and carry out two-dimensional scan, focus on the surface of sample 5 again by microcobjective 4, the photoacoustic signal that sample 5 is produced, survey by photoacoustic sensors 6, the output signal of photoacoustic sensors 6 is transported to capture card 8 and carries out data acquisition through after lock-in amplifier 7 amplifications and handling, and is stored and image reconstruction by computing machine 9 again.
The present invention compared with prior art has following advantage:
1, the beam flying technology that adopts no mechanical noise produces the photoacoustic signal with high spatial resolution in conjunction with high-resolution microcobjective, adopts photoacoustic sensors to carry out optoacoustic detection again, and pair cell carries out opto-acoustic microscopic imaging, and resolution is less than 1 micron.
2, adopt continuous laser to excite, need not adopt pulse laser to carry out photoacoustic imaging.
3, not only obtain the structural information of cell, but also the material composition of pair cell carries out imaging, obtain the function information of cell.
Description of drawings
Fig. 1 is a cell opto-acoustic microscopic imaging apparatus structure block diagram of the present invention;
Among the figure: the 1st, laser instrument, the 2nd, chopper, the 3rd, laser scanning galvanometer, the 4th, microcobjective, the 5th, sample, the 6th, photoacoustic sensors, the 7th, lock-in amplifier, the 8th, capture card, the 9th, computing machine;
Fig. 2 a is the optical microscopic image of erythrocyte;
Fig. 2 b is the optoacoustic micro-image of erythrocyte;
Fig. 2 c is the optical microscopic image of onion cell;
Fig. 2 d is the optoacoustic micro-image of onion cell.
Embodiment
As shown in Figure 1, apparatus of the present invention comprise laser scanning imaging mechanism, photoacoustic sensors and signal processor three parts.Wherein, laser scanning imaging mechanism is made of laser instrument 1, scanning galvanometer 3, microcobjective 4; During work, scanning galvanometer 3, microcobjective 4 and photoacoustic sensors 6 can be installed on the microscope stand;
Signal processor is made up of chopper 2, lock-in amplifier 7, capture card 8 and computing machine 9, and they are electrically connected successively.
The course of work of the present invention is as follows: the laser that laser instrument 1 sends, modulate by chopper 2, enter laser scanning galvanometer 3 then and carry out two-dimensional scan, focus on the surface of sample 5 again by microcobjective 4, the photoacoustic signal that sample 5 is produced is surveyed by photoacoustic sensors 6, and the output signal of photoacoustic sensors 6 is through after lock-in amplifier 7 amplifications and handling, be transported to capture card 8 and carry out data acquisition, store and image reconstruction by computing machine 9 again.Experimental result is compared with the optical microphotograph imaging shown in Fig. 2 a, Fig. 2 b, Fig. 2 c, Fig. 2 d, opto-acoustic microscopic imaging of the present invention have contain much information, the contrast advantages of higher, not only can obtain the structural information of cell, can also obtain the function information of cell.Cell outline is clear among the figure, can demonstrate the individual cells image.
Claims (3)
1. cell opto-acoustic microscopic imaging method is characterized in that:
---cell is placed on the concave surface of cover glass, laser sees through cover glass and is radiated on the cell, the cell absorbing light produces optoacoustic effect, be delivered to photoacoustic sensors by the sound pressure variations that this optoacoustic effect produced, survey and export photoacoustic signal then by photoacoustic sensors, to unicellular realization optoacoustic detection;
---by beam flying and high-resolution microcobjective, pair cell carries out two-dimensional scan, realizes single celled opto-acoustic microscopic imaging.
2. a cell opto-acoustic microscopic imaging device is characterized in that comprising laser scanning imaging mechanism, photoacoustic sensors and signal processor three parts; Wherein, laser scanning imaging mechanism is made of laser instrument, scanning galvanometer, microcobjective; Signal processor is electrically connected successively by chopper, lock-in amplifier, capture card and computing machine and constitutes.
3. according to the device shown in the claim 2, it is characterized in that the laser that laser instrument (1) sends, (2 modulate by chopper, enter scanning galvanometer (3) and carry out two-dimensional scan, focus on the surface of sample (5) again by microcobjective (4), the photoacoustic signal that sample (5) is produced, survey by photoacoustic sensors (6), the output signal of photoacoustic sensors (6) is through after lock-in amplifier (7) amplification and handling, be transported to capture card (8) and carry out data acquisition, store and image reconstruction by computing machine (9) again.
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201010115730 CN101782518A (en) | 2010-02-11 | 2010-02-11 | Cell opto-acoustic microscopic imaging method and device thereof |
| PCT/CN2010/073274 WO2011097851A1 (en) | 2010-02-11 | 2010-05-26 | Cell photo-acoustic microscope imaging method and device thereof |
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| CN 201010115730 CN101782518A (en) | 2010-02-11 | 2010-02-11 | Cell opto-acoustic microscopic imaging method and device thereof |
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| CN 201010115730 Pending CN101782518A (en) | 2010-02-11 | 2010-02-11 | Cell opto-acoustic microscopic imaging method and device thereof |
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102621067A (en) * | 2012-02-26 | 2012-08-01 | 曾吕明 | Real-time opto-acoustic imaging system based on two-dimensional laser galvanometer scanning |
| CN102621068A (en) * | 2012-02-26 | 2012-08-01 | 曾吕明 | Portable real-time opto-acoustic imaging system |
| CN102636434A (en) * | 2012-02-26 | 2012-08-15 | 曾吕明 | Portable three-dimensional visual photoacoustic imaging system |
| CN102636435A (en) * | 2012-02-26 | 2012-08-15 | 曾吕明 | Visible photo-acoustic imaging system based on three-dimensional dynamic focusing laser galvanometer scanning |
| CN103356174A (en) * | 2013-07-11 | 2013-10-23 | 贾宏博 | Device for high-speed two-photon microimaging and electrophysiology signal synchronous recording |
| CN103884690A (en) * | 2014-03-10 | 2014-06-25 | 湖南大学 | One-dimensional nano structure type three-dimensional direction microcell photovoltaic and lighting representing system and method |
| CN105212898A (en) * | 2015-08-31 | 2016-01-06 | 睿芯生命科技（深圳）有限公司 | A kind of reflective optoacoustic micro imaging system |
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| CN101336832A (en) * | 2008-08-12 | 2009-01-07 | 福建师范大学 | Pulse type optical acoustic scanning soft-tissue imaging method and device |
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- 2010-02-11 CN CN 201010115730 patent/CN101782518A/en active Pending
- 2010-05-26 WO PCT/CN2010/073274 patent/WO2011097851A1/en active Application Filing
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| WO2009055705A2 (en) * | 2007-10-25 | 2009-04-30 | Washington University In St. Louis | Confocal photoacoustic microscopy with optical lateral resolution |
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Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102621067A (en) * | 2012-02-26 | 2012-08-01 | 曾吕明 | Real-time opto-acoustic imaging system based on two-dimensional laser galvanometer scanning |
| CN102621068A (en) * | 2012-02-26 | 2012-08-01 | 曾吕明 | Portable real-time opto-acoustic imaging system |
| CN102636434A (en) * | 2012-02-26 | 2012-08-15 | 曾吕明 | Portable three-dimensional visual photoacoustic imaging system |
| CN102636435A (en) * | 2012-02-26 | 2012-08-15 | 曾吕明 | Visible photo-acoustic imaging system based on three-dimensional dynamic focusing laser galvanometer scanning |
| CN103356174A (en) * | 2013-07-11 | 2013-10-23 | 贾宏博 | Device for high-speed two-photon microimaging and electrophysiology signal synchronous recording |
| CN103356174B (en) * | 2013-07-11 | 2016-03-02 | 贾宏博 | The device of a kind of high speed two-photon micro-imaging and electro physiology synchronous recording |
| CN103884690A (en) * | 2014-03-10 | 2014-06-25 | 湖南大学 | One-dimensional nano structure type three-dimensional direction microcell photovoltaic and lighting representing system and method |
| CN105212898A (en) * | 2015-08-31 | 2016-01-06 | 睿芯生命科技（深圳）有限公司 | A kind of reflective optoacoustic micro imaging system |
| CN105212898B (en) * | 2015-08-31 | 2016-10-19 | 睿芯生命科技（深圳）有限公司 | A kind of reflective optoacoustic micro imaging system |
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| WO2011097851A1 (en) | 2011-08-18 |
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