CN106706577A - Optical imaging system and method - Google Patents
Optical imaging system and method Download PDFInfo
- Publication number
- CN106706577A CN106706577A CN201611022248.3A CN201611022248A CN106706577A CN 106706577 A CN106706577 A CN 106706577A CN 201611022248 A CN201611022248 A CN 201611022248A CN 106706577 A CN106706577 A CN 106706577A
- Authority
- CN
- China
- Prior art keywords
- light
- sample
- laser
- regional structure
- fluorescence signal
- 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
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/645—Specially adapted constructive features of fluorimeters
- G01N21/6456—Spatial resolved fluorescence measurements; Imaging
- G01N21/6458—Fluorescence microscopy
Landscapes
- Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Physics & Mathematics (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
- Microscoopes, Condenser (AREA)
Abstract
The invention discloses an optical imaging system and method, and is applicable to the technical field of optics. The system comprises a light source for generating laser, a spatial light modulator for modulating the laser to generate target stimulating laser and area structured light capable of addressing the target area, and illuminate a sample, generate fluorescence signals which carry cell change information after being stimulated by the target stimulating laser in the area. The system comprises a detector for recording the fluorescence signals, a controller for controlling the spatial light modulator to keep the target stimulating laser unchanged, moving a phase position of the area structured light to make the area structured light stripes on the sample move. The detector records the fluorescence signals generated after each time of moving the area structured light stripes. A control unit is arranged in the system for conducting spectrum analysis of the fluorescent images corresponding to all the recorded fluorescence signals to obtain a high resolution image of the target area under the influence of the target stimulating laser. The invention further discloses an optical imaging method, and can increase the imaging resolution of the structured lighting area.
Description
Technical field
The present invention relates to optical technical field, more particularly to a kind of photoimaging systems and method.
Background technology
Traditional fluorescence microscope has specific marker, can be to the advantage of target real time imagery, in life science
Serve extensive effect.However, traditional fluorescent microscopic imaging is influenceed by ssystem transfer function, limit system into
As resolution ratio, so as to limit effect of the fluorescent microscopic imaging in life science.In recent years, with a series of breakthrough diffraction poles
Limit, realizes the proposition of super-resolution imaging method, and super-resolution fluorescence technology turns into the focus of research work.For example：Stimulated emission is damaged
Consumption (STED, stimulated emission depletion) technology, Structured Illumination microscope (SIM, structured
Illumination microscopy) technology and based on unimolecule positioning super-resolution imaging technology --- photoactivation position
Microscope (PALM, photo-activation localization microscopy) and the aobvious mirror of random optical reconstruct
(STORM, stochastic optical reconstruction microscopy) etc..
Structured Illumination micro-imaging technique forms modulation image by changing the space structure of illumination light, in modulation image
Just contain the high-frequency information more than ssystem transfer function.Processed by calculating, extract the high-frequency information of modulation image, breached
Diffraction limit, reconstruct obtains the image of super-resolution.Structured Illumination micro-imaging technique can be applied directly on fluorescence microscope,
With simple structure, image taking speed is fast, and fluorescence molecule is not required particularly, can be applied to real time imagery, thus receives
It is extensive to pay attention to.
But current Structured Illumination micro-imaging technique, is to entirely regarding in addition to being illuminated to target area cell
The even whole sample in field carries out Structured Illumination, the energy of laser is wasted, while being caused to other cells outside target area
Photobleaching and phototoxicity.In addition, light channel structure is complicated, laser power requirement also limit greatly it and use scope.In research cell
Cell effect under the conditions of interphase interaction and environmental stimuli, and many cells the selection behavior of concurrent observation iuntercellular and structure
Relation, it is impossible to exclude the target under the conditions of the whole audience with the photic influence of noise of exterior domain.
The content of the invention
The present invention provides a kind of photoimaging methods system and method, is used to Laser Modulation into mesh by spatial light modulator
Mark stimulates light and regional structure light, is radiated at and fluorescence signal is produced on sample, records and analyze the fluorescence signal, can obtain the light
According to the high fdrequency component on direction, the resolution ratio of the illumination region direction can be improved, reduce photic influence of noise, save laser energy
Amount, and simplify light channel structure.
The present invention provides a kind of photoimaging methods system, including：
Light source, for producing laser；
Spatial light modulator, for being modulated to the laser, produces goal stimulus light and is addressed to sample object
The regional structure light in region, the goal stimulus light and the regional structure light irradiation on sample, in the target of the sample
Produced on region by the fluorescence signal that cellular change information is carried after the goal stimulus light stimulus；
Detector, for recording the fluorescence signal；
Control unit, for controlling the spatial light modulator to keep the goal stimulus light constant, the movement region
The phase of structure light so that the regional structure striations movement on the sample；
The detector, is additionally operable to record the fluorescence signal for moving produced after the regional structure striations every time；
Described control unit, is additionally operable to carry out spectrum analysis to the corresponding fluoroscopic image of all fluorescence signals for recording, and obtains
The high-definition picture of target area when must be influenceed by the goal stimulus light.
The present invention also provides a kind of photoimaging methods, including：
Produce laser；
Spatial light modulator is modulated to the laser, produces goal stimulus light and is addressed to sample object region
Regional structure light, the goal stimulus light and regional structure light irradiation are produced on sample on the target area of the sample
By the fluorescence signal that cellular change information is carried after the goal stimulus light stimulus；
Record the fluorescence signal；
Control spatial light modulator keeps the goal stimulus light constant, the phase of the movement regional structure light so that
Regional structure striations movement on the sample；
Record moves the fluorescence signal produced after the regional structure striations every time；
The corresponding fluoroscopic image of all fluorescence signals to recording carries out spectrum analysis, obtains and receives the goal stimulus shadow
The high-definition picture of target area when ringing.
Knowable to the embodiments of the present invention, by SLM by Laser Modulation into goal stimulus light and regional structure light, irradiate
On sample, the fluorescence signal of structural change of the cells is carried after target area life is by the goal stimulus light stimulus, and remember
The fluorescence signal is recorded, high fdrequency component can be obtained, you can improve the resolution ratio of the illumination region direction, control SLM keeps the thorn
Laser is constant, the phase of the mobile structure light so that the regional structure striations movement on sample, can improve mobile rear direction
Resolution ratio, record moves the fluorescence signal produced after the regional structure striations every time, and these fluorescence signals are carried out with frequency spectrum point
Analysis, the high-definition picture of target area when acquisition is influenceed by the goal stimulus light, after the move of stripe of whole plane is completed,
The imaging resolution of all directions illumination region in the plane can be improved, you can when obtaining stimulated light influence, specific region is thin
The high-definition picture of born of the same parents.
Brief description of the drawings
In order to illustrate more clearly about the embodiment of the present invention or technical scheme of the prior art, below will be to embodiment or existing
The accompanying drawing to be used needed for having technology description is briefly described, it should be apparent that, drawings in the following description are only this
Some embodiments of invention, for those skilled in the art, without having to pay creative labor, can be with root
Other accompanying drawings are obtained according to these accompanying drawings.
Fig. 1 is the light channel structure figure of photoimaging systems provided in an embodiment of the present invention；
Fig. 2 is that photoimaging methods provided in an embodiment of the present invention realize schematic flow sheet；
The schematic diagram of goal stimulus light and regional structure optical illumination mode in Fig. 3 (a)~3 (d) embodiment of the present invention.
Specific embodiment
To enable that goal of the invention of the invention, feature, advantage are more obvious and understandable, below in conjunction with the present invention
Accompanying drawing in embodiment, is clearly and completely described to the technical scheme in the embodiment of the present invention, it is clear that described reality
It is only a part of embodiment of the invention to apply example, and not all embodiments.Based on the embodiment in the present invention, people in the art
The every other embodiment that member is obtained under the premise of creative work is not made, belongs to the scope of protection of the invention.
A kind of photoimaging systems and method by one-photon excitation structure light are the embodiment of the invention provides, be can be used for
The nano-resolution structural information of living cells is studied under the conditions of the light stimulus of any-mode with the response of light stimulus.The present embodiment profit
One or more goal stimuluses Guang Yu areas needed for being formed with spatial light modulator (SLM, Spatial Light Modulator)
Domain structure optical illumination pattern, is radiated in the specific region of microscopical sample, excites generation fluorescence to believe based on single photon effect
Number, and be imaged on detector (EMCCD, Electron-Multiplying CCD) by microcobjective collection.Can be according to plan
Be stimulated the stimulation sites and imaging region of target, at the same produce stimulate light (luminous point) and be stimulated object matching regional structure
Light, it can be one, or multiple points to stimulate luminous point, and the position of point can arbitrarily be adjusted as needed.Imageable target can
To be one, or multiple, can arbitrarily select as needed.When being stimulated light stimulus shadow by surrounding target cell
Ring, the fluoroscopic image is contained stimulates high-frequency information of the corresponding illumination region cell at moment more than ssystem transfer function.Control
SLM moves different structure light, obtains corresponding modulation image, and calculating can be obtained by goal stimulus light stimulus time domain or cell
High lateral resolution image, can therefrom analyze the super-resolution knot of the cell under cell interaction and incentive condition
Structure information.It is further explained below.
Fig. 1 is referred to, Fig. 1 is a kind of light channel structure schematic diagram of photoimaging systems.The photoimaging systems include：
Light source 1, for producing laser.
Specifically, light source 1 can use visible laser, such as blue laser, and the wavelength of the blue laser can be with
It is 405nm (nanometer), or 488nm.Such as ultraviolet laser again, the wavelength of the ultraviolet laser can be
350nm.Can be used for the laser or mercury lamp and ultraviolet LD (Laser Diode, semiconductor laser) of single photon fluorescence imaging
With LED can, the present embodiment relax to light source selection requirement so that the optional face of light source is wider.
Spatial light modulator (SLM) 5, for being modulated to the laser that light source 1 is produced, produces goal stimulus light and can seek
Location to the target area of sample regional structure light, the goal stimulus light and regional structure light irradiation on sample 15, in sample
Produced on 15 target area by the fluorescence signal after the goal stimulus light stimulus, the cell of the target area is subject to the target
After stimulation, eucaryotic cell structure is changed, and cellular change information is carried in the fluorescence signal for producing.The spatial light modulator
It can be reflective pure phase type spatial light modulator.The phase of incident light is modulated using SLM5, needed for reflecting
Goal stimulus light and regional structure light pattern.
Sample 15 is placed under microscopical microcobjective 14, in the present embodiment, eucaryotic cell structure is included in sample 15.
The goal stimulus light, specific region or specific cells for stimulating sample 15, the regional structure light, for illuminating
The cell of specific region around the stimulation light, on sample 15, generation is pierced by this for the goal stimulus light and regional structure light irradiation
The specific region carries the fluorescence signal of cellular change information during laser effect.
Detector 18, for recording the fluorescence signal.
Control unit 19, can be a control module in computer, or computer, in Fig. 1 by taking computer as an example.Control
Unit processed 19 is connected with SLM5, and controllable SLM5 produces the goal stimulus light and regional structure light, also can control SLM5 and keeps
The goal stimulus light is constant, the phase of the mobile regional structure light so that the regional structure striations movement on sample 15.Control
Unit 19 can control SLM5 and keep stimulating light constant, and moving area structural light stripes in one direction,
Detector 18, is additionally operable to record the fluorescence signal for moving produced after the regional structure striations every time.
Regional structure striations is often mobile once, the record first order fluorescence signal of detector 18.Moving structure striations is imaged,
The resolution ratio for rotating rear direction can be improved.The like, after the move of stripe in a target area is completed, it is possible to
Improve the imaging resolution of the illumination zone in the plane in all directions target area.
The fluorescence signal of the record of detector 18 is stored in control unit 19.
Control unit 19, is additionally operable to carry out spectrum analysis to the corresponding fluoroscopic image of all fluorescence signals for recording, and obtains
The high-definition picture of target area when being influenceed by the goal stimulus light.
All fluorescence signals of the record of detector 18 all form fluoroscopic image, and detector 18 is connected with control unit 19, will
The fluoroscopic image of record is passed in control unit 19, and control unit 19 carries out frequency spectrum point to these fluoroscopic images according to Predistribution Algorithm
Analysis, when acquisition is influenceed by goal stimulus light, the high-definition picture in target area.
Further, also it is sequentially provided between light source 1 and SLM5：Beam-expanding collimation system and a half-wave plate.Wherein,
The beam-expanding collimation system includes two lens, i.e. lens 2 and lens 3, and the beam-expanding collimation system is used for the laser for sending light source 1
Expand the collimated light beam to form preset size.Half-wave plate 4, the polarization state for changing the collimated light beam after beam-expanding collimation, changes
It is the linearly polarized light for polarizing in the target direction.
Further, the photoimaging systems also include：Spatial filter.
The spatial filter includes two lens and a diaphragm, and two lens are lens 6 and lens 8, and a diaphragm is
Diaphragm 7.Two lens constitute 4f systems, the rear focus of the first lens 6 in two lens and the front focus weight of the second lens 8
Close, diaphragm 7 is located in the focus that two lens overlap, for will end by the zero order diffracted light after SLM5.Can carry
The signal to noise ratio of imaging is risen, effectively using the dynamic range of detector.
Further, the photoimaging systems also include：Fourier lense 9, speculum 10, pipe mirror 11.
Wherein, fourier lense 9, for will be focused on the mirror 10 by the light after the spatial filter；
Speculum 10, for the light that fourier lense 9 is focused on to be reflexed into pipe mirror 11；
Pipe mirror 11, for the light that speculum 10 reflects to be formed into parallel hot spot.
Further, the photoimaging systems also include：Exciter filter 12, dichroic mirror 13, microscope.
Microscope is entered by exciter filter 12, dichroic mirror 13 from the parallel hot spot out of pipe mirror 11, it is micro- at this
Under the focussing force of the microcobjective 14 of mirror, formation goal stimulus light and regional structure on the focal plane of microcobjective 14 is focused on
Light, the focal plane of microcobjective 14 overlaps with the sample face of sample 15.
Further, the photoimaging systems also include：Transmitting optical filter 16, the 3rd lens 17；
Transmitting optical filter 16 is bandpass filter, for high anti-to laser, fluorescence signal is turned on, and only allows fluorescence signal
Pass through.
3rd lens 17, fluorescence signal produced on the target area of sample 15 for that will pass through focuses on detection
On device 18.
In the present embodiment, by SLM by Laser Modulation into goal stimulus light and regional structure light, it is radiated on sample,
Target area generation is received to carry the fluorescence signal of cellular change information after the goal stimulus light stimulus, and records fluorescence letter
Number, high fdrequency component can be obtained, you can improve the resolution ratio of the illumination region direction, control SLM keeps the stimulation light constant, moves
Move the phase of the regional structure light so that the regional structure striations movement on sample, the resolution ratio of mobile rear direction can be improved,
Record moves the fluorescence signal produced after the regional structure striations every time, and these fluorescence signals are carried out with spectrum analysis, obtains
The high-definition picture of target area when being influenceed by the goal stimulus light, after the move of stripe of whole plane is completed, can improve
The imaging resolution of all directions illumination region in the plane, you can when obtaining stimulated light influence, the height of specific region cell
Image in different resolution.Meanwhile, light channel structure is simplified, it is cost-effective.
Refer to Fig. 2, Fig. 2 is that a kind of photoimaging methods realize schematic flow sheet.The photoimaging methods include：
S201, generation laser；
Laser is produced by light source.The light source be can single photon fluorescence imaging laser.
S202, spatial light modulator are modulated to the laser, produce goal stimulus light and are addressed to sample object area
The regional structure light in domain, the goal stimulus light and regional structure light irradiation are produced on sample on the target area of the sample
By the fluorescence signal that cellular change information is carried after the goal stimulus light stimulus；
SLM produces goal stimulus light and is addressed to the regional structure light in sample object region to Laser Modulation, is radiated at
On sample, fluorescence signal is produced in the target area of sample, the fluorescence signal is that goal stimulus light is subject on target area
The fluorescence signal produced after stimulation, after the cell of the target area is subject to the goal stimulus, eucaryotic cell structure changes, and is producing
The fluorescence signal in carry cellular change information.
Specifically, including two beam-expanding collimation systems of lens carry out the laser to expand the directional light to form preset size
Beam, half-wave plate will expand after the collimated light beam the polarization state linear polarization that is changed to polarize in the target direction by half-wave plate
Light, reflective pure phase type spatial light modulator is modulated to the phase for changing the laser after polarization state, produces initial mesh
Mark stimulates light and regional structure light.
After carrying out space filtering, focusing, reflection successively to the initial goal stimulus light and regional structure light, by Guan Jing
Form parallel hot spot.Wherein, space filtering is realized by spatial filter, and the spatial filter includes two lens and one
Individual diaphragm, two lens constitute 4f systems, the rear focus of the first lens in two lens and the front focus weight of the second lens
Close, the relation of the first lens and the second lens before and after forming position in light path, diaphragm is located at the focus that two lens overlap
On, for will end by the zero order diffracted light after SLM.Afterwards, by fourier lense focus the light on the mirror,
Goal stimulus light and regional structure light pattern are formed on the minute surface of speculum, speculum reflexes to the light that fourier lense is focused on
On Guan Jing, parallel hot spot is formd by Guan Jing.
The parallel hot spot sequentially passes through exciter filter, dichroic mirror, microcobjective, gathers on the focal plane of the microcobjective
Jiao forms illumination and excites hot spot, i.e. the goal stimulus light and regional structure light.The goal stimulus light and regional structure light irradiation exist
On sample, produce by the fluorescence that cellular change information is carried after the goal stimulus light stimulus on the target area of the sample
Signal.
S203, record the fluorescence signal；
The fluorescence signal sequentially passes through dichroic mirror, transmitting optical filter, lens after being collected by the microcobjective, by detector
Receive and record.
Transmitting optical filter only allows fluorescence signal to pass through.
Angle between dichroic mirror and incident light can be 45 degree.
S204, control spatial light modulator keep the goal stimulus light constant, the phase of the mobile regional structure light so that
Regional structure striations movement on the sample；
SLM and detector are respectively connected with computer, and computer can control SLM to produce by the control unit of its own
Raw the goal stimulus light and regional structure light, also can control SLM and keep the goal stimulus light constant, the mobile regional structure light
Phase so that the regional structure striations movement on sample.The controllable SLM of control unit keeps stimulating light constant, and towards one
Direction moving area structural light stripes.
S205, record move the fluorescence signal produced after the regional structure striations every time；
Regional structure striations is often mobile once, and detector just records first order fluorescence signal.Moving structure striations is imaged,
The resolution ratio for rotating rear direction can be improved.The like, after the move of stripe in a target area is completed, it is possible to
Improve the imaging resolution of the illumination zone in the plane in all directions target area.
S206, the corresponding fluoroscopic image of all fluorescence signals to recording carry out spectrum analysis, obtain and receive the goal stimulus
The high-definition picture of target area when light influences.
All fluorescence signals of detector record all form fluoroscopic image, and the fluoroscopic image of record is passed to calculating by detector
In machine, control unit carries out spectrum analysis to these fluoroscopic images according to Predistribution Algorithm, when acquisition receives the influence of goal stimulus light,
High-definition picture in target area.
In one example, Fig. 3 (a)~Fig. 3 (d) is referred to, Fig. 3 (a)~Fig. 3 (d) is target thorn in the embodiment of the present invention
The schematic diagram of laser and regional structure optical illumination mode, specifically, Fig. 3 (a) is unicellular by single stimulation light, many cells matching
Excitation structure light, have in figure a cell be subject to one stimulation light luminous point stimulate, Fig. 3 (b) be it is unicellular by thorniness laser,
Many cells match excitation structure light, have a cell to be subject to 4 luminous point stimulations of stimulation light in figure, and Fig. 3 (c) is that many cells are subject to
It is single to stimulate light, many cells to match excitation structure light, there are 3 cells each to be stimulated by the luminous point of 1 stimulation light respectively in figure；Figure
3 (d) is that many cells are subject to thorniness laser, many cells to match excitation structure light, has 3 cells to be each subject to 2,3,4 respectively in figure
The individual luminous point for stimulating light stimulates.In Fig. 3 (a)~Fig. 3 (d), structure light illuminates multiple cells, and structure light illuminates scope not
Become.
In the present embodiment, by SLM by Laser Modulation into goal stimulus light and regional structure light, it is radiated on sample,
Target area life is recorded the fluorescence signal by the fluorescence signal after the goal stimulus light stimulus, can obtain high fdrequency component,
The resolution ratio of the illumination region direction can be improved, control SLM keeps the stimulation light constant, the phase of the mobile structure light makes
The regional structure striations movement on sample is obtained, the resolution ratio of mobile rear direction can be improved, record moves the regional structure every time
These fluorescence signals are carried out spectrum analysis, mesh when acquisition is influenceed by the goal stimulus light by the fluorescence signal produced after striations
The high-definition picture in region is marked, after the move of stripe of whole plane is completed, all directions lighting area in the plane can be improved
The imaging resolution in domain, you can when obtaining stimulated light influence, the high-definition picture of specific region cell.
In the above-described embodiments, the description to each embodiment all emphasizes particularly on different fields, and does not have the portion described in detail in certain embodiment
Point, may refer to the associated description of other embodiments.
It is more than the description to photoimaging systems provided by the present invention and method, for those skilled in the art, according to
According to the thought of the embodiment of the present invention, will change in specific embodiments and applications, to sum up, in this specification
Appearance should not be construed as limiting the invention.
Claims (9)
1. a kind of photoimaging systems, it is characterised in that the system includes：
Light source, for producing laser；
Spatial light modulator, for being modulated to the laser, produces goal stimulus light and is addressed to sample object region
Regional structure light, the goal stimulus light and the regional structure light irradiation on sample, in the target area of the sample
It is upper to produce by the fluorescence signal that cellular change information is carried after the goal stimulus light stimulus；
Detector, for recording the fluorescence signal；
Control unit, for controlling the spatial light modulator to keep the goal stimulus light constant, the movement regional structure
The phase of light so that the regional structure striations movement on the sample；
The detector, is additionally operable to record the fluorescence signal for moving produced after the regional structure striations every time；
Described control unit, is additionally operable to carry out spectrum analysis to the corresponding fluoroscopic image of all fluorescence signals for recording, and is received
The high-definition picture of target area when the goal stimulus light influences.
2. system according to claim 1, it is characterised in that between the light source and the spatial light modulator also successively
It is provided with：Beam-expanding collimation system and a half-wave plate；
The beam-expanding collimation system includes two lens, and the laser beam expanding for the light source to be sent forms the flat of preset size
Row light beam；
The half-wave plate, the polarization state for the collimated light beam after by beam-expanding collimation is changed to the line for polarizing in the target direction
Polarised light.
3. system according to claim 1 and 2, it is characterised in that the system also includes：Spatial filter；
The spatial filter includes two lens and a diaphragm, and described two lens constitute 4f systems, described two lens
In the rear focus of the first lens overlapped with the front focus of the second lens, the diaphragm is located at the focus that described two lens overlap
On, for will end by the zero order diffracted light after the spatial light modulator.
4. system according to claim 3, it is characterised in that the system also includes：Fourier lense, speculum, pipe
Mirror；
The fourier lense, for the speculum will to be focused on by the light after the spatial filter；
The speculum, for the light that the fourier lense is focused on to be reflexed into the Guan Jing；
The Guan Jing, for the light that the speculum reflects to be formed into parallel hot spot.
5. system according to claim 4, it is characterised in that the system also includes：It is exciter filter, dichroic mirror, aobvious
Micro mirror；
The parallel hot spot enters microscope by the exciter filter, dichroic mirror, in the microscopical microcobjective
Under focussing force, the goal stimulus light and regional structure light, Jiao are formed on the focal plane for focusing on the microcobjective
Plane overlaps with the sample face of the sample.
6. system according to claim 5, it is characterised in that the system also includes：Transmitting optical filter, the 3rd lens；
The transmitting optical filter is bandpass filter, for high anti-to laser, fluorescence signal is turned on；
3rd lens, the fluorescence signal produced on the target area of the sample for that will pass through focuses on institute
State on detector.
7. a kind of photoimaging methods, it is characterised in that methods described includes：
Produce laser；
Spatial light modulator is modulated to the laser, produces goal stimulus light and is addressed to the region in sample object region
Structure light, the goal stimulus light and regional structure light irradiation are produced on the target area of the sample and receive institute on sample
State after goal stimulus light stimulus and carry the fluorescence signal of cellular change information；
Record the fluorescence signal；
Control spatial light modulator keeps the goal stimulus light constant, the phase of the movement regional structure light so that described
Regional structure striations movement on sample；
Record moves the fluorescence signal produced after the regional structure striations every time；
The corresponding fluoroscopic image of all fluorescence signals to recording carries out spectrum analysis, when acquisition is influenceed by the goal stimulus light
The high-definition picture of target area.
8. method according to claim 7, it is characterised in that the spatial light modulator is modulated to the laser,
Produce goal stimulus light and be addressed to the regional structure light in sample object region, the goal stimulus light and regional structure illumination
Penetrate on sample, produce on the target area of the sample and carried cellular change by after the goal stimulus light stimulus
The fluorescence signal of information includes：
The laser is carried out to expand the collimated light beam to form preset size；
The linearly polarized light that the polarization state of the collimated light beam after expanding is changed to polarize in the target direction by half-wave plate；
Reflective pure phase type spatial light modulator is modulated to the phase for changing the laser after polarization state, produces initial mesh
Mark stimulates light and regional structure light；
After carrying out space filtering, focusing, reflection successively to the initial goal stimulus light and regional structure light, by pipe mirror shape
Into parallel hot spot；
The parallel hot spot sequentially passes through exciter filter, dichroic mirror, microcobjective, gathers on the focal plane of the microcobjective
Jiao forms the goal stimulus light and regional structure light；
The goal stimulus light and regional structure light irradiation are produced on the target area of the sample and receive the mesh on sample
The fluorescence signal of cellular change information is carried after mark stimulation light stimulus.
9. the method according to claim 7 or 8, it is characterised in that described to record the fluorescence signal and include：
The fluorescence signal sequentially passes through dichroic mirror, transmitting optical filter, lens after being collected by the microcobjective, by detector
Receive and record.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611022248.3A CN106706577B (en) | 2016-11-16 | 2016-11-16 | A kind of photoimaging systems and method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611022248.3A CN106706577B (en) | 2016-11-16 | 2016-11-16 | A kind of photoimaging systems and method |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN106706577A true CN106706577A (en) | 2017-05-24 |
| CN106706577B CN106706577B (en) | 2019-08-16 |
Family
ID=58941060
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201611022248.3A Active CN106706577B (en) | 2016-11-16 | 2016-11-16 | A kind of photoimaging systems and method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106706577B (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107024850A (en) * | 2017-05-26 | 2017-08-08 | 清华大学 | High-speed structures light 3-D imaging system |
| CN107272218A (en) * | 2017-05-26 | 2017-10-20 | 清华大学 | High-speed structures photoimaging systems |
| CN111024671A (en) * | 2019-12-31 | 2020-04-17 | 深圳大学 | System and method for performing super-resolution imaging on directional photostimulation structural change |
| CN111060485A (en) * | 2019-12-31 | 2020-04-24 | 苏州栢科昇科技有限公司 | Microorganism multi-modal imaging system and microorganism multi-modal imaging detection method |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101655601A (en) * | 2008-08-22 | 2010-02-24 | 麦克奥迪实业集团有限公司 | Light microscope imaging method and system based on DMD structure |
| WO2014186544A1 (en) * | 2013-05-15 | 2014-11-20 | The Administrators Of The Tulane Educational Fund | Microscopy of a tissue sample using structured illumination |
| WO2015033514A1 (en) * | 2013-09-06 | 2015-03-12 | 株式会社ニコン | Illumination device and structured illumination microscope |
| CN104515759A (en) * | 2014-12-16 | 2015-04-15 | 中国科学院苏州生物医学工程技术研究所 | Non-linear structure light illumination microscopic imaging method and system |
| CN105784653A (en) * | 2016-03-08 | 2016-07-20 | 浙江大学 | Wide-field super resolution fluorescence microscopic imaging device |
| WO2016151666A1 (en) * | 2015-03-20 | 2016-09-29 | 株式会社ニコン | Structured illumination microscope, observation method, and image processing program |
-
2016
- 2016-11-16 CN CN201611022248.3A patent/CN106706577B/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101655601A (en) * | 2008-08-22 | 2010-02-24 | 麦克奥迪实业集团有限公司 | Light microscope imaging method and system based on DMD structure |
| WO2014186544A1 (en) * | 2013-05-15 | 2014-11-20 | The Administrators Of The Tulane Educational Fund | Microscopy of a tissue sample using structured illumination |
| WO2015033514A1 (en) * | 2013-09-06 | 2015-03-12 | 株式会社ニコン | Illumination device and structured illumination microscope |
| CN104515759A (en) * | 2014-12-16 | 2015-04-15 | 中国科学院苏州生物医学工程技术研究所 | Non-linear structure light illumination microscopic imaging method and system |
| WO2016151666A1 (en) * | 2015-03-20 | 2016-09-29 | 株式会社ニコン | Structured illumination microscope, observation method, and image processing program |
| CN105784653A (en) * | 2016-03-08 | 2016-07-20 | 浙江大学 | Wide-field super resolution fluorescence microscopic imaging device |
Non-Patent Citations (3)
| Title |
|---|
| MING ZHAO ET AL: "Cellular imaging of deep organ using two-photon Bessel light-sheet nonlinear structured illumination microscopy", 《BIOMEDICAL OPTICS EXPRESS》 * |
| R. AYUK ET AL: "Structured illumination fluorescence microscopy with distorted excitations using a filtered blind-SIM algorithm", 《OPTICS LETTERS》 * |
| 吴美瑞 等: "结构光照明荧光显微镜突破衍射极限的原理和在生命科学中的应用", 《激光与光电子学进展》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107024850A (en) * | 2017-05-26 | 2017-08-08 | 清华大学 | High-speed structures light 3-D imaging system |
| CN107272218A (en) * | 2017-05-26 | 2017-10-20 | 清华大学 | High-speed structures photoimaging systems |
| CN111024671A (en) * | 2019-12-31 | 2020-04-17 | 深圳大学 | System and method for performing super-resolution imaging on directional photostimulation structural change |
| CN111060485A (en) * | 2019-12-31 | 2020-04-24 | 苏州栢科昇科技有限公司 | Microorganism multi-modal imaging system and microorganism multi-modal imaging detection method |
Also Published As
| Publication number | Publication date |
|---|---|
| CN106706577B (en) | 2019-08-16 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN107167929B (en) | Double mode optical ultra-discrimination microscopic imaging device and method based on DMD | |
| CN107389631A (en) | The multi-modal Structured Illumination super-resolution micro imaging system of high speed polychrome and its method | |
| CN104515759A (en) | Non-linear structure light illumination microscopic imaging method and system | |
| CN106706577B (en) | A kind of photoimaging systems and method | |
| CN106133580B (en) | Stimulated emission depletion microscopie unit | |
| US8097864B2 (en) | System, method and computer-accessible medium for providing wide-field superresolution microscopy | |
| Ducros et al. | Encoded multisite two-photon microscopy | |
| CN110954523B (en) | Two-photon scanning structure light microscopic imaging method and device | |
| CN111982870B (en) | Scanning structure light super-resolution microscopic imaging device and method | |
| CN103487421B (en) | Time gate wide field stimulated radiation super-resolution microscopic method and device | |
| CN110954522A (en) | Multifocal super-resolution optical microscopic imaging method and device | |
| CN110082900A (en) | Variable illumination Fourier overlapping associations imaging device, system and method | |
| CN106094189B (en) | The compound microscopic system of multichannel fluorescence microscopy | |
| CN102540446A (en) | High-speed structure illumination optical microscope system and method based on digital micromirror device | |
| CN108181235B (en) | STED parallel microscopic imaging system based on uniform structured light illumination | |
| CN103605202A (en) | Structured light illumination microscopic imaging system based on silicon-based liquid crystal chip | |
| CN103744172B (en) | A kind of confocal microscopic image method possessing spatial light modulation illumination | |
| CN108196357A (en) | A kind of multi-angle illumination light source and the Fourier stacking imaging system based on this light source | |
| CN110320654A (en) | Based on the microscopical quick three-dimensional body imaging system of multi-angle 4Pi and method | |
| CN114965405B (en) | Super-resolution single-objective light sheet microscopic imaging system | |
| CN110824681A (en) | Non-scanning high super-resolution optical three-dimensional microscopic imaging method | |
| CN108897126B (en) | Fluorescence imaging system | |
| CN113946044A (en) | Multi-focus multi-photon microscopic imaging system and method based on point spread function engineering | |
| Kozawa et al. | Light needle microscopy with spatially transposed detection for axially resolved volumetric imaging | |
| CN111024671B (en) | System and method for super-resolution imaging of directional light stimulation structural change |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |