CN108318464A - A kind of super-resolution fluorescence fluctuation micro imaging method, device and storage medium - Google Patents
A kind of super-resolution fluorescence fluctuation micro imaging method, device and storage medium Download PDFInfo
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Abstract
The invention discloses a kind of super-resolution fluorescence fluctuation micro imaging method, device and storage medium, the super-resolution fluorescence fluctuation micro imaging method is several width detection images that sample of the acquisition after fluorescent molecular marks passes through fluorescence microimaging systems；Fourier transformation is carried out to the detection image, obtains the Fourier spectrum of several width detection images；The initial estimate of sample structure and fluorescence intensity fluctuations is set, and the initial estimate based on the sample structure and fluorescence intensity fluctuations and several width detection images carry out default time exporting the imaging results of the sample after iterating to calculate according to default iterative algorithm.By regarding the Strength Changes of scintillation fluor as random speckle needed for the imaging of ordinary construction light, and super resolution image is obtained by iterative algorithm, both it had not needed additional experimental provision and has introduced structure light, also the resolution ratio that Structured Illumination micro-imaging can be improved under less detection times, the balance for realizing resolution ratio and being imaged between cost.
Description
Technical field
The present invention relates to micro-imaging technique field, more particularly to a kind of super-resolution fluorescence fluctuation micro imaging method, dress
It sets and storage medium.
Background technology
The one kind of fluorescent optics microtechnic as light microscope, spatial resolution are similarly subjected to the limit of diffraction limit
System.In order to break through diffraction limit, current existing method includes that Structured Illumination microtechnic (SIM), unimolecule positioning are micro-
(STORM, PALM) and fluorescence fluctuation imaging (SOFI) etc..Wherein SIM is using the structure light irradiating sample of design, to system
Point spread function be modulated, to obtain the high-frequency information of sample, realize super-resolution imaging.But this method is introducing
When structure light, it is necessary to some additional devices, while the highest frequency of structure light is limited by diffraction limit, which has limited
Accessible highest resolution ratio in common SIM imaging theories.
And STORM, PALM and SOFI are realized using special natures such as switching effect, the scintillation effects of fluorescent molecular
The method of super-resolution imaging.Wherein, STORM and PALM belongs to unimolecule positioning microtechnic, is to utilize fluorescence photoswitch effect
Or other mechanism Random sparseness excite fluorescent molecular, then to the method that sparse hot spot is positioned, a nanometer fraction can be reached
Resolution.And SOFI is then to utilize scintillation fluor effect, the method for statistical analysis to the sequence of intensity changed over time.This is several
The kind required detection times of method are far longer than SIM imagings, and the temporal resolution that this allows for this method is relatively low.
Thus the prior art could be improved and improve.
Invention content
Place in view of above-mentioned deficiencies of the prior art, the purpose of the present invention is to provide a kind of fluctuation of super-resolution fluorescence is micro-
Imaging method, device and storage medium, by by the Strength Changes of scintillation fluor be used as ordinary construction light imaging needed for
Machine speckle, and obtain super resolution image by iterative algorithm, had not both needed additional experimental provision and has introduced structure light, also can compared with
The resolution ratio that Structured Illumination micro-imaging is improved under few detection times realizes flat between resolution ratio and imaging cost
Weighing apparatus.
In order to achieve the above object, this invention takes following technical schemes：
A kind of super-resolution fluorescence fluctuation micro imaging method comprising following steps：
Acquire several width detection images that the sample after fluorescent molecular marks passes through fluorescence microimaging systems；
Fourier transformation is carried out to the detection image, obtains the Fourier spectrum of several width detection images；
The initial estimate of sample structure and fluorescence intensity fluctuations is set, the sample structure and fluorescence intensity fluctuations are based on
Initial estimate and several width detection images Fourier spectrum, default iterative calculation is carried out according to default iterative algorithm
The imaging results of the sample are exported afterwards.
In the super-resolution fluorescence fluctuation micro imaging method, sample of the acquisition after fluorescent molecular marks passes through
In the step of several width detection images of fluorescence microimaging systems, several width detection images are：Wherein N is the quantity of detection image, and S (r) is sample structure, Pn(r)(n
=1 ..., N) it is fluorescence intensity fluctuations, PSF (r) is the point spread function of fluorescence microimaging systems.
It is described that Fourier transformation is carried out to the detection image in the super-resolution fluorescence fluctuation micro imaging method,
In the step of obtaining the Fourier spectrum of several width detection images, the Fourier spectrum of several width detection images is：Wherein, On=SPn,WithRespectively InAnd OnFourier spectrum, OTF is fluorescence
The optical transfer function of micro imaging system.
In the described super-resolution fluorescence fluctuation micro imaging method, the setting sample structure and fluorescence intensity fluctuations just
Beginning estimated value, the Fourier of initial estimate and several width detection images based on the sample structure and fluorescence intensity fluctuations
Frequency spectrum, carrying out the step of exporting the imaging results of the sample after default iterative calculation according to default iterative algorithm includes：
Sample structure S and fluorescence intensity fluctuations P is setn(n=1 ..., N) it changes to n-th of the wherein primary of detection image
Initial estimate during generation is respectively SgAnd Pn,g(n=1 ..., N)；
According to SgAnd Pn,gCalculate the fluorescence distribution and its Fourier spectrum of n-th of detection image；
The Fourier spectrum of the fluorescence distribution is updated according to n-th of detection image, and passes through Fourier's inversion
Change the fluorescence distribution for obtaining updated n-th of detection image；
According to the fluorescence distribution of n-th of detection image and updated n-th of detection image to SgAnd Pn,gIt is updated,
Obtain updated value Sg updateAnd Pn,g update；
Judge whether current n is less than N, if so, enabling n=n+1, returns to the fluorescence for continuing to calculate (n+1)th detection image
Distribution and its Fourier spectrum, until N number of detection image is fully completed iteration, by SgAnd Pn,g(n=1 ..., N) all it is updated to
Sg updateAnd Pn,g update, (n=1 ..., N)；
By Sg updateAnd Pn,g update, n=1 ..., N replace SgAnd Pn,g, n=1 ..., N, be re-used as call by value return after
The continuous fluorescence distribution and its Fourier spectrum for calculating (n+1)th detection image will export result until reaching default iterations
SgImaging results as the sample.
It is described according to S in the super-resolution fluorescence fluctuation micro imaging methodgAnd Pn,gCalculate n-th detection image
The step of fluorescence distribution and its Fourier spectrum includes：
According to SgAnd Pn,gCalculate the fluorescence distribution O of n-th of detection imagen,g=Sg·Pn,g；
Fourier transformation is carried out to the fluorescence distribution of n-th of detection image, obtains the fluorescence point of n-th of detection image
The Fourier spectrum of cloth
It is described that the fluorescence is divided according to n-th of detection image in the super-resolution fluorescence fluctuation micro imaging method
The Fourier spectrum of cloth is updated, and the fluorescence distribution of updated n-th of detection image is obtained by inverse Fourier transform
The step of include：
The Fourier spectrum of the fluorescence distribution is updated according to n-th of detection image, obtains updated fluorescence
The Fourier spectrum of publication
Inverse Fourier transform is carried out to the Fourier spectrum of updated fluorescence distribution, obtains updated n-th of detection
The fluorescence distribution of image
It is described according to n-th of detection image and updated n-th in the super-resolution fluorescence fluctuation micro imaging method
The fluorescence distribution of a detection image is to SgAnd Pn,gIt is updated, obtains updated value Sg updateAnd Pn,g updateThe step of include：
Based on equationWith
P respectivelyn,gTo SgBe updated, obtain updated value Sg updateAnd Pn,g update, wherein | | indicate absolute value.
A kind of super-resolution fluorescence fluctuation microscopic imaging device comprising：Processor, memory and communication bus；
The computer-readable program that can be executed by the processor is stored on the memory；
The communication bus realizes the connection communication between processor and memory；
The processor realized when executing the computer-readable program super-resolution fluorescence fluctuation as described above it is micro- at
Step in image space method.
A kind of computer readable storage medium, which is characterized in that the computer-readable recording medium storage there are one or
The multiple programs of person, one or more of programs can be executed by one or more processor, as described above super to realize
Resolved fluorometric fluctuates the step in micro imaging method.
Compared to the prior art, super-resolution fluorescence fluctuation micro imaging method, device and storage medium provided by the invention
In, super-resolution fluorescence fluctuation micro imaging method by acquire the sample after fluorescent molecular marks by fluorescence microscopy at
As several width detection images of system；Fourier transformation is carried out to the detection image, obtains several width detection images
Fourier spectrum；The initial estimate of sample structure and fluorescence intensity fluctuations is set, the sample structure and fluorescence intensity are based on
The Fourier spectrum of the initial estimate of fluctuation and several width detection images carries out default iteration according to default iterative algorithm
The imaging results of the sample are exported after calculating.Needed for the Strength Changes of scintillation fluor are used as in the imaging of ordinary construction light
Random speckle, and super resolution image is obtained by iterative algorithm, had not both needed additional experimental provision and introduced structure light, and also could
The resolution ratio that Structured Illumination micro-imaging is improved under less detection times realizes between resolution ratio and imaging cost
Balance.
Description of the drawings
Fig. 1 is the flow chart that super-resolution fluorescence provided by the invention fluctuates micro imaging method.
Fig. 2 is the structure diagram that super-resolution fluorescence provided by the invention fluctuates microscopic imaging device.
Fig. 3 is iterative process schematic diagram in Application Example of the present invention.
Fig. 4 is the super-resolution imaging schematic diagram of existing wide field imaging.
Fig. 5 is the imaging signal that the super-resolution fluorescence that first preferred embodiment of the invention provides fluctuates micro imaging method
Figure.
Fig. 6 is the imaging signal that the super-resolution fluorescence that second preferred embodiment of the invention provides fluctuates micro imaging method
Figure.
Fig. 7 be fig. 4 to fig. 6 in along 1 normalized intensity scatter chart.
Fig. 8 be fig. 4 to fig. 6 in along 2 normalized intensity scatter chart.
Specific implementation mode
It can not meet time and spatial resolution simultaneously in view of fluorescent microscopic imaging in the prior art and low cost requires
The shortcomings of, the purpose of the present invention is to provide super-resolution fluorescence fluctuation micro imaging method, device and storage medium, by will be glimmering
The Strength Changes of optical flare pass through iterative algorithm and obtain super-resolution figure as random speckle needed for the imaging of ordinary construction light
Picture had not both needed additional experimental provision and has introduced structure light, it is micro- also to improve Structured Illumination under less detection times
The resolution ratio of imaging, the balance for realizing resolution ratio and being imaged between cost.
To make the purpose of the present invention, technical solution and effect clearer, clear and definite, develop simultaneously embodiment pair referring to the drawings
The present invention is further described.It should be appreciated that described herein, specific examples are only used to explain the present invention, is not used to
Limit the present invention.
Referring to Fig. 1, super-resolution fluorescence fluctuation micro imaging method provided by the invention includes the following steps：
Several width detection images that the sample of S100, acquisition after fluorescent molecular marks passes through fluorescence microimaging systems；
S200, Fourier transformation is carried out to the detection image, obtains the Fourier spectrum of several width detection images；
S300, the initial estimate that sample structure and fluorescence intensity fluctuations is arranged are strong based on the sample structure and fluorescence
The initial estimate of fluctuation and several width detection images are spent, is exported after carrying out default iterative calculation according to default iterative algorithm
The imaging results of the sample.
The present invention carries out fluorescent marker to sample first, utilizes scintillation fluor characteristic or fluorescent switch effect, acquisition warp
Sample after fluorescent molecular label is by several width detection images of fluorescence microimaging systems, i.e., a series of fluorescence intensities are at any time
Between the detection image that changes, Fourier transformation is carried out to the detection image later, obtains the Fourier of several width detection images
The initial estimate of sample structure and fluorescence intensity fluctuations is arranged in frequency spectrum later, is based on the sample structure and fluorescence intensity wave
The Fourier spectrum of dynamic initial estimate and several width detection images carries out default iteration meter according to default iterative algorithm
The imaging results of the sample are exported after calculation, i.e. the present invention is used as by the Strength Changes of scintillation fluor in the imaging of ordinary construction light
Required random speckle, and super resolution image is obtained by iterative algorithm, it had not both needed additional experimental provision and has introduced structure light,
Also the resolution ratio that Structured Illumination micro-imaging can be improved under less detection times, realize resolution ratio with imaging cost it
Between balance.
Specifically, in the step S100, several width detection image sequences are expressed as：
Wherein N is the quantity of detection image, and S (r) is sample structure, Pn(r) (n=1 ..., N) is fluorescence intensity fluctuations,
A series of fluorescence intensity fluctuations that i.e. scintillation fluor generates, PSF (r) are the point spread function of fluorescence microimaging systems.
In the step S200, Fourier transformation is carried out to the detection image, i.e., in a frequency domain, formula (1) is expressed as：
Wherein, On=SPn,WithRespectively InAnd OnFourier spectrum, OTF is fluorescence microimaging systems
Optical transfer function, InIt is it is known that S and P with OTFnTo be unknown, S is then subsequently reconstructed by iterative algorithm, obtains and is ultimately imaged
As a result.
Specifically, the step S300 includes：
S301, setting sample structure S and fluorescence intensity fluctuations Pn(n=1 ..., N) to n-th of detection image wherein
Initial estimate during an iteration is respectively SgAnd Pn,g(n=1 ..., N)；
S302, according to SgAnd Pn,gCalculate the fluorescence distribution and its Fourier spectrum of n-th of detection image；
S303, the Fourier spectrum of the fluorescence distribution is updated according to n-th of detection image, and passes through Fourier
Inverse transformation obtains the fluorescence distribution of updated n-th of detection image；
S304, according to the fluorescence distribution of n-th of detection image and updated n-th of detection image to SgAnd Pn,gIt carries out
Update obtains updated value Sg updateAnd Pn,g update；
S305, judge whether current n is less than N, if so, enabling n=n+1, return and continue (n+1)th detection image of calculating
Fluorescence distribution and its Fourier spectrum, until N number of detection image is fully completed iteration, by SgAnd Pn,g(n=1 ..., N) whole is more
It is newly Sg updateAnd Pn,g update, (n=1 ..., N)；
S306, by Sg updateAnd Pn,g update, n=1 ..., N replace SgAnd Pn,g, n=1 ..., N are re-used as call by value and return
Returning the fluorescence distribution and its Fourier spectrum for continuing to calculate (n+1)th detection image will export until reaching default iterations
As a result SgImaging results as the sample.
In the present embodiment, sample structure S and fluorescence intensity fluctuations P is first setn(n=1 ..., N) to n-th of detection figure
Initial estimate during the wherein an iteration of picture is respectively SgAnd Pn,g(n=1 ..., N), starts iterative process later,
The fluorescence distribution and its Fourier spectrum of n-th of detection image, the specific step S302 packets are first calculated according to initial estimate
It includes：
S3021, according to SgAnd Pn,gCalculate the fluorescence distribution O of n-th of detection imagen,g=Sg·Pn,g；
S3022, Fourier transformation is carried out to the fluorescence distribution of n-th of detection image, n-th of detection image of acquisition
The Fourier spectrum of fluorescence distribution
N-th of detection image I is utilized laternThe Fourier spectrum of the fluorescence distribution is updated, and it is carried out
Inverse Fourier transform obtains the fluorescence distribution of updated n-th of detection image, and the specific step S303 includes：
S3031, the Fourier spectrum of the fluorescence distribution is updated according to n-th of detection image, after being updated
Fluorescence publication Fourier spectrum
S3032, inverse Fourier transform is carried out to the Fourier spectrum of updated fluorescence distribution, obtains updated n-th
The fluorescence distribution of a detection image
Later according to the fluorescence distribution of n-th of detection image and updated n-th of detection image to SgAnd Pn,gInto
Row update, obtains updated value Sg updateAnd Pn,g update, specific renewal process is based on equationWith
P respectivelyn,gTo SgBe updated, obtain updated value Sg updateAnd Pn,g update, wherein | | indicate absolute value；Obtain updated value
Updated value S is used afterwardsg updateInstead of current Sg, judge whether current n is less than N, if so, enable n=n+1, return continue calculating n-th+
The fluorescence distribution and its Fourier spectrum of 1 detection image, and utilize In+1To calculate Pn+1,gUpdated value, until N number of detection scheme
As being fully completed iteration, i.e. n=N, by SgAnd Pn,g(n=1 ..., N) all it is updated to Sg updateAnd Pn,g update, (n=1 ...,
N)；At this time by Sg updateAnd Pn,g update, n=1 ..., N replace SgAnd Pn,g, n=1 ..., N are re-used as call by value and return and continue
The fluorescence distribution and its Fourier spectrum of (n+1)th detection image of calculating, the Fourier spectrum update of the above-mentioned fluorescence distribution of repetition,
SgAnd Pn,gRenewal process, until reaching default iterations, by final output SgAs the imaging results of the sample,
And then realize super-resolution imaging, due to being not necessarily to introduce structure light, there is no need to additional experimental provisions, and are positioned with unimolecule
Microtechnic is compared, and the intensity number that the present invention generates scintillation fluor is used as random speckle, and is obtained and surpassed by iterative algorithm
Resolution image, needed for detection times be significantly less than unimolecule positioning microtechnic, that is, meet time and spatial resolution
Requirement, also reduce the cost of super-resolution imaging system.
Micro imaging method is fluctuated based on the super-resolution fluorescence that above-described embodiment provides, the present invention correspondingly provides a kind of meter
Calculation machine readable storage medium storing program for executing, there are one either multiple programs are one or more for the computer-readable recording medium storage
A program can be executed by one or more processor, to realize in super-resolution fluorescence fluctuation micro imaging method as described above
The step of.It is described in detail due to fluctuating micro imaging method to the super-resolution fluorescence above, is not described further herein,
Referring specifically to the corresponding embodiment of the above method.
Micro imaging method is fluctuated based on the super-resolution fluorescence that above-described embodiment provides, the present invention correspondingly provides a kind of super
Resolved fluorometric fluctuates microscopic imaging device, as shown in Fig. 2, super-resolution fluorescence fluctuation microscopic imaging device processor 10, with
The memory 20 that the processor 10 communicates to connect, and for the communication interface 30 of signal transmission, the processor 10, storage
Device 20 and communication interface 30 are connected by communication bus 40.Wherein, the memory 20 is stored with computer-readable program,
The computer-readable program is executed by processor 10 to realize that super-resolution fluorescence as described above fluctuates micro imaging method；Institute
Processor 10 is stated for calling the computer program in the memory 20, is shown with executing super-resolution fluorescence fluctuation as described above
Micro- imaging method.Since super-resolution fluorescence fluctuation micro imaging method being described in detail above, do not make herein
It is described in detail, referring specifically to the corresponding embodiment of the above method.
The iterative process of micro imaging method is fluctuated to super-resolution fluorescence provided by the invention below in conjunction with specific embodiment
And the image reconstruction result of super-resolution fluorescence fluctuation micro imaging method illustrates：
1, fluorescence probe and sample of the selection with blinking characteristic carries out fluorescent marker to sample；
2, suitable experimental provision and systematic parameter are selected in conjunction with the characteristic of sample, including optical source wavelength, detector are big
Small, microscopic system numerical aperture, enlargement ratio, sample frequency, sampling number etc.；
3, sample is sampled using the existing fluorescence microimaging systems put up, obtains a series of Strength Changes
Image；
4, the initial estimate for selecting sample structure and fluorescence intensity fluctuations, is updated in iterative algorithm and is iterated, full
The image exported after sufficient condition is just used as and is ultimately imaged result.
Specifically, as shown in figure 3, super-resolution fluorescence provided by the invention fluctuates the middle iterative algorithm of micro imaging method
Iterative process is：
(1) according to estimated value SgAnd Pn,gIt is O to calculate n-th of fluorescence distributionn,g=Sg·Pn,g, frequency spectrum
(2) n-th of detection image I is utilizednTo frequency spectrumIt is updated：
(3) it can be obtained by by inverse Fourier transform
(4) utilize iterative algorithm to SgAnd Pn,gIt is updated, while in view of fluorescence excitation light field is positive value, therefore
Newer formula is as follows：
Wherein | | indicate absolute value.
(5) S is usedg updateInstead of Sg, step (1) is returned to, enables n=n+1, and utilize In+1To calculate Pn+1,gUpdated value, directly
It is fully completed iteration to N number of detection image, at this time former estimated value SgAnd Pn,g(n=1 ..., N) all it is updated to Sg updateWith
Pn,g update, (n=1 ..., N).
(6) by Sg updateAnd Pn,g update, n=1 ..., N replace SgAnd Pn,g, n=1 ..., N are re-used as call by value, heavy
The multiple above process presets iterations until meeting, terminates iteration.
In order to verify the feasibility that super-resolution fluorescence provided by the invention fluctuates micro-imaging (hereinafter referred to as SRFB) method
And resolution ratio, as shown in Fig. 4 to Fig. 8, the imaging results being imaged with existing wide field to SRFB methods illustrate, wherein testing
Parameter is as follows：Wavelength of fluorescence is 690nm, and frame frequency 25fps, detection image size is 128pixel × 128pixel, detector
Valid pixel size is 100nm, and NA=1.3, detection image quantity is 500.In order to improve excitation concentration to reduce required visit
Number is surveyed, every 10 width of 500 width images can be superimposed together to obtain the image of 50 width highers excitation concentration, it can also be by every 20
Width is superimposed together to obtain 25 width.For both of these case, Fig. 4, Fig. 5 and Fig. 6 be respectively wide-field image (widefield),
SRFB handle the super-resolution imaging that 25 width images (srfb, N=25) and 50 width images (srfb, N=50) obtain as a result, Fig. 7 with
Fig. 8 be fig. 4 to fig. 6 in respectively along 1 and line 2 normalized intensity distribution curve.The halfwidth for calculating Gaussian Profile in Fig. 8 can
Halfwidth to obtain single micro-tubular structure in fig. 4 to fig. 6 is respectively 393nm, 276nm and 229nm, this explanation works as detection image
When quantity is 50, image that super-resolution fluorescence provided by the invention fluctuation micro imaging method obtains, can be with compared to wide field imaging
Realize that being more than 2 times of resolution ratio improves, and without increasing additional experimental provision.
In conclusion in super-resolution fluorescence fluctuation micro imaging method provided by the invention, device and storage medium, it is described
Super-resolution fluorescence fluctuates micro imaging method and passes through fluorescence microimaging systems by acquiring the sample after fluorescent molecular marks
Several width detection images；Fourier transformation is carried out to the detection image, obtains the Fourier of several width detection images
Frequency spectrum；The initial estimate of sample structure and fluorescence intensity fluctuations is set, based on the sample structure and fluorescence intensity fluctuations
The Fourier spectrum of initial estimate and several width detection images, after carrying out default iterative calculation according to default iterative algorithm
Export the imaging results of the sample.It is random needed for the imaging of ordinary construction light by being used as the Strength Changes of scintillation fluor
Speckle, and super resolution image is obtained by iterative algorithm, it had not both needed additional experimental provision and has introduced structure light, it also can be less
Detection times under improve Structured Illumination micro-imaging resolution ratio, realize resolution ratio and be imaged cost between balance.
It, can according to the technique and scheme of the present invention and its hair it is understood that for those of ordinary skills
Bright design is subject to equivalent substitution or change, and all these changes or replacement should all belong to the guarantor of appended claims of the invention
Protect range.
Claims (9)
1. a kind of super-resolution fluorescence fluctuates micro imaging method, which is characterized in that include the following steps：
Acquire several width detection images that the sample after fluorescent molecular marks passes through fluorescence microimaging systems；
Fourier transformation is carried out to the detection image, obtains the Fourier spectrum of several width detection images；
The initial estimate of sample structure and fluorescence intensity fluctuations is set, based on the sample structure and fluorescence intensity fluctuations just
The Fourier spectrum of beginning estimated value and several width detection images carries out default time defeated after iterating to calculate according to default iterative algorithm
Go out the imaging results of the sample.
2. super-resolution fluorescence according to claim 1 fluctuates micro imaging method, which is characterized in that the acquisition is through fluorescence
Sample after molecular labeling is by the step of several width detection images of fluorescence microimaging systems, several width detection figures
As being： Wherein N is the quantity of detection image, and S (r) is sample structure, Pn
(r) (n=1 ..., N) is fluorescence intensity fluctuations, and PSF (r) is the point spread function of fluorescence microimaging systems.
3. super-resolution fluorescence according to claim 2 fluctuates micro imaging method, which is characterized in that described to the detection
In the step of image progress Fourier transformation, the Fourier spectrum of acquisition several width detection images, several width detections
The Fourier spectrum of image is： Wherein, On=SPn,WithRespectively InAnd OnFu
In leaf frequency spectrum, OTF be fluorescence microimaging systems optical transfer function.
4. super-resolution fluorescence according to claim 3 fluctuates micro imaging method, which is characterized in that the setting sample knot
The initial estimate of structure and fluorescence intensity fluctuations, if the initial estimate based on the sample structure and fluorescence intensity fluctuations and
The Fourier spectrum of dry width detection image, according to default iterative algorithm export after default iterative calculation the sample at
As the step of result includes：
Sample structure S and fluorescence intensity fluctuations P is setn(n=1 ..., N) in the wherein an iteration mistake to n-th of detection image
Initial estimate in journey is respectively SgAnd Pn,g(n=1 ..., N)；
According to SgAnd Pn,gCalculate the fluorescence distribution and its Fourier spectrum of n-th of detection image；
The Fourier spectrum of the fluorescence distribution is updated according to n-th of detection image, and is obtained by inverse Fourier transform
Obtain the fluorescence distribution of updated n-th of detection image；
According to the fluorescence distribution of n-th of detection image and updated n-th of detection image to SgAnd Pn,gIt is updated, obtains more
New value Sg updateAnd Pn,g update；
Judge whether current n is less than N, if so, enabling n=n+1, returns to the fluorescence distribution for continuing to calculate (n+1)th detection image
And its Fourier spectrum, until N number of detection image is fully completed iteration, by SgAnd Pn,g(n=1 ..., N) all it is updated to
Sg updateAnd Pn,g update, (n=1 ..., N)；
By Sg updateAnd Pn,g update, n=1 ..., N replace SgAnd Pn,g, n=1 ..., N are re-used as call by value return and continue to count
The fluorescence distribution and its Fourier spectrum for calculating (n+1)th detection image will export result S until reaching default iterationsgMake
For the imaging results of the sample.
5. super-resolution fluorescence according to claim 4 fluctuates micro imaging method, which is characterized in that described according to SgAnd Pn,g
The step of fluorescence distribution and its Fourier spectrum for calculating n-th of detection image includes：
According to SgAnd Pn,gCalculate the fluorescence distribution O of n-th of detection imagen,g=Sg·Pn,g；
Fourier transformation is carried out to the fluorescence distribution of n-th of detection image, obtains the fluorescence distribution of n-th of detection image
Fourier spectrum
6. super-resolution fluorescence according to claim 5 fluctuates micro imaging method, which is characterized in that described according to n-th
Detection image is updated the Fourier spectrum of the fluorescence distribution, and obtains updated n-th by inverse Fourier transform
The step of fluorescence distribution of a detection image includes：
The Fourier spectrum of the fluorescence distribution is updated according to n-th of detection image, obtains updated fluorescence publication
Fourier spectrum
Inverse Fourier transform is carried out to the Fourier spectrum of updated fluorescence distribution, obtains updated n-th of detection image
Fluorescence distribution
7. super-resolution fluorescence according to claim 6 fluctuates micro imaging method, which is characterized in that described according to n-th
The fluorescence distribution of detection image and updated n-th of detection image is to SgAnd Pn,gIt is updated, obtains updated value Sg updateWith
Pn,g updateThe step of include：
Based on equationWith
P respectivelyn,gTo SgBe updated, obtain updated value Sg updateAnd Pn,g update, wherein | | indicate absolute value.
8. a kind of super-resolution fluorescence fluctuates microscopic imaging device, which is characterized in that including：Processor, memory and communication bus；
The computer-readable program that can be executed by the processor is stored on the memory；
The communication bus realizes the connection communication between processor and memory；
The processor realizes that the super-resolution as described in claim 1-7 any one is glimmering when executing the computer-readable program
Step in light beats micro imaging method.
9. a kind of computer readable storage medium, which is characterized in that the computer-readable recording medium storage there are one or
Multiple programs, one or more of programs can be executed by one or more processor, to realize that claim 1-7 such as appoints
The step in super-resolution fluorescence fluctuation micro imaging method described in meaning one.
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| CN113670865A (en) * | 2020-05-13 | 2021-11-19 | 深圳华大智造科技有限公司 | Resolution board, resolution evaluation method and related equipment |
| CN113973167A (en) * | 2021-10-28 | 2022-01-25 | 维沃移动通信有限公司 | Camera assembly, electronic device and image generation method |
| CN116740705A (en) * | 2023-04-06 | 2023-09-12 | 深圳大学 | Super-bright light spot positioning and identifying method based on random forest classification algorithm |
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| CN113973167A (en) * | 2021-10-28 | 2022-01-25 | 维沃移动通信有限公司 | Camera assembly, electronic device and image generation method |
| WO2024065094A1 (en) * | 2022-09-26 | 2024-04-04 | 广州超视计生物科技有限公司 | Super-resolution microscopic imaging method and apparatus, and computer device and storage medium |
| CN116740705A (en) * | 2023-04-06 | 2023-09-12 | 深圳大学 | Super-bright light spot positioning and identifying method based on random forest classification algorithm |
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