CN105319725A - Ultra-high resolution imaging method used for rapid moving object - Google Patents
Ultra-high resolution imaging method used for rapid moving object Download PDFInfo
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- 238000003384 imaging method Methods 0.000 title claims abstract description 139
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- 238000012800 visualization Methods 0.000 abstract 1
- 230000003287 optical effect Effects 0.000 description 25
- 230000008569 process Effects 0.000 description 8
- 238000004458 analytical method Methods 0.000 description 3
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- G—PHYSICS
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- G02B27/00—Optical systems or apparatus not provided for by any of the groups G02B1/00 - G02B26/00, G02B30/00
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- G06V10/25—Determination of region of interest [ROI] or a volume of interest [VOI]
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- G06T—IMAGE DATA PROCESSING OR GENERATION, IN GENERAL
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Abstract
The invention relates to an ultra-high resolution imaging method used for a rapid moving object, comprising: according to the position of an ultra-high resolution imaging target on a moving object sample, setting an ROI including the ultra-high resolution imaging target; obtaining the moving speed of the ultra-high resolution imaging target and the moving speed and position of the ROI, and obtaining the imaging frame frequency of an ultra-high resolution imaging module; calculating the size of an imaging shooting area Sp according to the relation between the imaging frame frequency and the Sp; defining a pre-visualization area Si including the ultra-high resolution imaging target on the sample, wherein the Si has the same size with the Sp, and the same moving speed with the ROI; adjusting the position of the Si or the Sp according to the motion of the ROI so as to keep the Si and the Sp overlapped; and according to the Si, performing ultra-high resolution imaging on the ultra-high resolution imaging target on the moving object sample. The ultra-high resolution imaging method can eliminate the influence of object motion to ultra-high resolution imaging.
Description
Technical field
The present invention relates to acquisition technology field, particularly relate to a kind of super-resolution formation method for fast moving objects.
Background technology
Generally, the size of the discernmible smallest object of human eye is approximately 0.1mm.See less object if want, then need by means of microtechnic.1873, German microtechnic expert Ernest & Whitney. Abbe discloses optical microscope due to the diffraction effect of light and finite aperture resolution and there is the principle of the limit, this principle " constraint " utilization of conventional optical microscope at Nanometer World just.
When only having single fluorescence molecule under the microscopical object lens visual field, by specific algorithm matching, can easily exceed optical resolution limit.For exploring microworld, the super-resolution microtechnic breaking through the optical limit of optical microscope is arisen at the historic moment.Within 1981, first unimolecule tracking technique is incorporated in life science by Barak and Webb.Although monomolecular registration can reach nanoscale, it can not improve the resolution of light microscope when differentiating two or more pointolite.
Within 2002, Patterson and Lippincott-Schwartz utilizes the mutation of green fluorescent protein (GFP) (PA-GFP) to observe specified protein at intracellular movement locus first.Germany EricBezig recognizes observantly: application single molecular fluorescence imaging technique, in conjunction with the characteristics of luminescence of this fluorescin, can break through the limit of optical resolution---photoactivation location microtechnic (PALM) has been born.The formation method of PALM can only be used for observing heterogenous expression albumen, helpless to cellular endogenous albumen.2006, U.S. Howard-Hughes research institute scientist Zhuan Xiao of Chinese origin common vetch experimental group found: different wavelength can control the switching of chemiluminescence molecule Cy5 between fluorescence excitation state and dark-state.Given this random optical reconstruct microtechnic (STORM) is developed.No matter be PALM or STORM super-resolution microscopy, its point spread function imaging is still consistent with traditional micro-imaging, needs repeatedly to activate-quenching fluorescence molecule, so experiment completes mostly on fixed cells.
2000, Germany scientist StefanHell proposed the spot size being reduced exciting light by physical process, and the halfwidth directly reducing point spread function improves resolution, successfully have developed stimulated emission depletion microtechnic (STED).The another kind of method changing point spread function realization breakthrough optical diffraction limit is saturated structures illumination microtechnic (SSIM).2005, first nonlinear organization illumination optical parts were incorporated on traditional microscope by Gustafsson, obtained the image that resolution reaches 50nm.But existing super-resolution techniques image taking speed is slow, and be difficult to the sample taking motion (particularly rapid movement).
Summary of the invention
One of them object of the present invention is to provide a kind of super-resolution formation method for fast moving objects, to solve the technical matters that image taking speed in prior art is difficult to take moving object, particularly biological living slowly.
For achieving the above object, embodiments provide a kind of super-resolution formation method for fast moving objects, comprising:
Position according to super-resolution imageable target on moving object sample arranges the region of interest ROI comprising described super-resolution imageable target;
Obtain the movement velocity V of described super-resolution imageable target
iwith the movement velocity V of described region of interest ROI
rwith position;
According to the resolution PPI of super-resolution image-forming module
s, described region of interest ROI movement velocity V
rand the movement velocity V of described super-resolution imageable target
iobtain the imaging frame frequency f of described super-resolution image-forming module
s;
According to described imaging frame frequency f
sthe size of described imaging shooting area Sp is calculated with the relation of imaging shooting area Sp; Sample defines the plan shooting area Si that comprises super-resolution imaging target, the size of described plan shooting area Si and the measure-alike of described imaging shooting area Sp, and with region of interest ROI, there is identical movement velocity V
r;
Adjust the position of described plan shooting area Si or described imaging shooting area Sp, keep overlapping with described imaging shooting area Sp to make described plan shooting area Si; And according to described plan shooting area Si, super-resolution imaging is carried out to super-resolution imageable target on moving object sample.
Alternatively, described in
According to the resolution PPI of super-resolution image-forming module
s, described region of interest ROI movement velocity V
rand the movement velocity V of described super-resolution imageable target
iobtain the imaging frame frequency f of described super-resolution image-forming module
sstep in adopt following formula to obtain described imaging frame frequency f
s:
|V
r-V
i|/PPI
s≤f
s。
Alternatively, the movement velocity V of the described super-resolution imageable target of described acquisition
iwith the movement velocity V of region of interest ROI
rwith position by obtaining with under type:
At least two images of region of interest ROI are gathered respectively in Preset Time;
The position of moving object sample and super-resolution imageable target at least two images described in contrast, to obtain the displacement of moving object sample and super-resolution imageable target;
The movement velocity V of described region of interest ROI is obtained according to the displacement of moving object sample and super-resolution imageable target and imaging frame frequency
rand the movement velocity V of described super-resolution imageable target
i.
Alternatively, the movement velocity V of the described super-resolution imageable target of described acquisition
iwith the movement velocity V of region of interest ROI
robtain in the following manner with position:
In region of interest ROI selected one with first representative point that can represent its motion feature, super-resolution imageable target is selected the second representative point that can represent its motion feature;
The light intensity of described first representative point and described second representative point is gathered respectively in the different moment;
The variation relation of the light intensity time of the first representative point described in comparative analysis and described second representative point, to obtain displacement and the speed of moving object sample area-of-interest and super-resolution imageable target.
Alternatively, the position of described adjustment described plan shooting area Si or described imaging shooting area Sp, to make described plan shooting area Si keep in the step overlapped with described imaging shooting area Sp, realize described plan shooting area Si in the following manner and keep overlapping with described imaging shooting area Sp:
According to the movement velocity V of described region of interest ROI
radjust the position of described imaging shooting area Sp, make that the motion of described imaging shooting area Sp is identical with described region of interest ROI direction of motion, velocity magnitude is equal.
Alternatively, the position of described adjustment described plan shooting area Si or described imaging shooting area Sp, to make described plan shooting area Si keep in the step overlapped with described imaging shooting area Sp, realize described plan shooting area Si in the following manner and keep overlapping with described imaging shooting area Sp:
Described imaging shooting area Sp remains unchanged, according to the movement velocity V of described region of interest ROI
radjustment sample cell, produces the movement velocity V with described region of interest ROI to make sample
rdirection of motion is contrary, the equal-V of velocity magnitude
r.
The embodiment of the present invention is by obtaining position and the movement velocity of moving object sample area-of-interest and super-resolution imageable target, and in conjunction with the resolution determination super-resolution imaging speed of super-resolution image-forming module, take this as a foundation and be adjusted to the area of picture shooting area, ensure super-resolution imaging rate, thus the impact that elimination photographic subjects is taken for super-resolution relative to the motion of ROI; By being adjusted to the position as the position of shooting area or sample being intended shooting area, to make imaging shooting area keep overlapping with the position of sample being intended shooting area, thus the motion eliminating moving object sample ROI is on the impact of super-resolution imaging.The present invention can carry out super-resolution imaging to fast moving objects quickly and automatically, also can the morphosis of automatic analysis moving object, is particularly useful for the rapid ultra high resolution imaging of the various Moving Objects such as spermatoblast, biological tissue.
Accompanying drawing explanation
Can understanding the features and advantages of the present invention clearly by reference to accompanying drawing, accompanying drawing is schematic and should not be construed as and carry out any restriction to the present invention, in the accompanying drawings:
Fig. 1 is the schematic flow sheet of a kind of super-resolution formation method for fast moving objects that one embodiment of the invention provides;
Fig. 2 is a kind of super-resolution image device structure schematic diagram for fast moving objects that one embodiment of the invention provides;
Fig. 3 is adjusted to picture shooting area to adjust with plan shooting area the process schematic that overlaps in one embodiment of the invention.
Embodiment
Below in conjunction with drawings and Examples, the specific embodiment of the present invention is described in further detail.Following examples for illustration of the present invention, but are not used for limiting the scope of the invention.
First aspect, embodiments provides a kind of super-resolution formation method for fast moving objects, as shown in Figure 1, comprising:
Position according to super-resolution imageable target on moving object sample arranges the region of interest ROI comprising described super-resolution imageable target;
Obtain the movement velocity V of described super-resolution imageable target
iwith the movement velocity V of described region of interest ROI
rwith position;
According to the resolution PPI of super-resolution image-forming module
s, described region of interest ROI movement velocity V
rand the movement velocity V of described super-resolution imageable target
iobtain the imaging frame frequency f of described super-resolution image-forming module
s;
According to described imaging frame frequency f
sthe size of described imaging shooting area Sp is calculated with the relation of imaging shooting area Sp; Definition one comprises the plan shooting area Si of super-resolution imaging target, the size of described plan shooting area Si and the measure-alike of described imaging shooting area Sp, and has identical movement velocity V
r;
Adjust the position of described plan shooting area Si or described imaging shooting area Sp, keep overlapping with described imaging shooting area Sp to make described plan shooting area Si; And according to described plan shooting area Si, super-resolution imaging is carried out to super-resolution imageable target on moving object sample.
The embodiment of the present invention by obtaining position and the movement velocity of moving object sample and super-resolution imageable target, and obtains the area of imaging shooting area in conjunction with the resolution of super-resolution image-forming module, ensure super-resolution imaging rate; Keep overlapping to make the position of intending shooting area with imaging shooting area by the position be adjusted to as shooting area or sample, thus the motion of elimination moving object sample is on the impact of super-resolution imaging.The present invention can carry out super-resolution imaging to moving object quickly and automatically, also can the morphosis of automatic analysis moving object, is particularly useful for the rapid ultra high resolution imaging of the various Moving Objects such as spermatoblast, biological tissue.
Alternatively, according to the resolution PPI of super-resolution image-forming module
s, described region of interest ROI movement velocity V
rand the movement velocity V of described super-resolution imageable target
iobtain the imaging frame frequency f of described super-resolution image-forming module
sstep in adopt following formula obtain imaging frame frequency f
s:
|V
r-V
i|/PPI
s≤f
s。
Alternatively, the movement velocity V of described super-resolution imageable target is obtained
iwith the movement velocity V of region of interest ROI
robtain in the following manner with position:
Obtain the position of moving object sample at least two images and super-resolution imageable target;
At least two images of region of interest ROI are gathered respectively in Preset Time;
Contrast the position of moving object sample and super-resolution imageable target at least two images, to obtain the displacement of moving object sample and super-resolution imageable target;
The movement velocity V of described region of interest ROI is obtained according to the displacement of moving object sample and super-resolution imageable target and imaging frame frequency
rand the movement velocity V of described super-resolution imageable target
i.
Alternatively, the movement velocity V of region of interest ROI is obtained
robtain in the following manner with position:
In region of interest ROI selected one with first representative point that can represent its motion feature, super-resolution imageable target is selected the second representative point that can represent its motion feature;
The light intensity of the first representative point and the second representative point is gathered respectively in the different moment;
The variation relation of the light intensity time of comparative analysis first representative point and the second representative point, to obtain displacement and the speed of moving object sample region of interest ROI and super-resolution imageable target.
Will be understood that, above " its motion feature can be represented " and refer to, the first representative point is identical with the direction of motion of region of interest ROI, velocity magnitude is equal.In practical application, obtained movement velocity and the position of this point by position sensor.
In practical application, for making imaging shooting area Sp and region of interest ROI keep overlapping, realize in the following manner:
According to the movement velocity V of region of interest ROI
rbe adjusted to the motion feature of picture shooting area Sp, make that imaging shooting area Sp is identical with region of interest ROI direction of motion, velocity magnitude is equal.Or imaging shooting area Sp remains unchanged, according to the movement velocity V of region of interest ROI
radjustment sample cell, produces the movement velocity-V contrary with described region of interest ROI direction of motion, velocity magnitude is equal to make sample
r.
For embodying the superiority of a kind of super-resolution formation method for fast moving objects that the embodiment of the present invention provides, the embodiment of the present invention additionally provides a kind of super-resolution imaging side device for fast moving objects, as shown in Figure 2, comprising:
Conventional resolution micro-imaging module 10, for showing moving object sample and super-resolution imageable target to provide position and the movable information of this moving object sample and super-resolution imageable target;
Rapid image Acquire and process module 20, for obtaining the movement velocity V of moving object sample, the movement velocity V of super-resolution imageable target
i, intend the movement velocity V of shooting area Si
rwith the size of position and imaging shooting area Sp, and be supplied to position feedback control module 40 and super-resolution image-forming module 30;
Position feedback control module 40, for adjusting the position of described imaging shooting area Sp or plan imaging region Si, keeps overlapping with imaging shooting area Sp to make intending shooting area Si;
Super-resolution image-forming module 30, for according to super-resolution imaging rate requirement, is adjusted to the size of picture shooting area Sp, carries out super-resolution imaging to super-resolution imageable target on moving object sample.
As a kind of concrete example of conventional resolution micro-imaging module 10, as shown in Figure 2, the conventional resolution micro-imaging module 10 that the embodiment of the present invention provides comprises lighting unit and image-generating unit.Lighting unit comprises the light fixture 101 be successively set on the second imaging optical path, the 3rd lens 102 and condenser 103, and wherein, the side of the 3rd lens 102 is provided with light fixture 101, and opposite side is provided with condenser 103; The side away from light fixture 101 of this condenser 103 is provided with Electrocontrolled sample platform 104.Image-generating unit comprises imaging len 107, optical filtering 108 on the image-forming objective lens 105 be successively set on the second imaging optical path, optical splitter 106 and the 3rd imaging optical path; The side of image-forming objective lens 105 is provided with Electrocontrolled sample platform 104, and opposite side is provided with optical splitter 106; The light that 106 optical splitters change on the second imaging optical path forms the 3rd imaging optical path; Imaging len 107 is arranged on the side of optical splitter 106, and opposite side is provided with optical filtering 108.
In practical application, those skilled in the art can according to concrete use scenes, and select the conventional resolution micro-imaging module of suitable parameters to realize display fast moving objects sample and super-resolution imageable target, the present invention is not construed as limiting.
As a kind of concrete example of rapid image Acquire and process module 20, as shown in Figure 2, the rapid image Acquire and process module 20 that the embodiment of the present invention provides comprises rapid image collecting device 201 and computing equipment 202, wherein:
Rapid image collecting device 201, for gathering moving object sample and super-resolution imageable target to provide position and the movable information of this moving object sample and super-resolution imageable target;
Computing equipment 202, for according to the position of moving object sample and super-resolution imageable target and movable information to obtain the movement velocity V of moving object sample, the movement velocity V of super-resolution imageable target
i, intend the movement velocity V of shooting area Si
rwith the size of position and imaging shooting area Sp.
In practical application, rapid image collecting device 201 can adopt such as charge coupled cell (Charge-coupledDevice, or complementary metal oxide semiconductor (CMOS) (ComplementaryMetalOxideSemiconductor CCD), the imageing sensor such as CMOS), also can use location sensing detector etc., those skilled in the art can select according to concrete use scenes, and the present invention is not construed as limiting.
In practical application, computing equipment 202 is according to the similarity etc. of the ground unrest at least two images, moving object sample A morphosis, moving object sample A and background, take the technique computes movement velocitys such as different background modelings, target identification, Track Pick-up, those skilled in the art can select suitable disposal route according to concrete use scenes.
As a kind of concrete example of position feedback control module 40, as shown in Figure 2, the position feedback control module 40 that the embodiment of the present invention provides comprises the first lens 403, second lens 402 and the automatically controlled tilting mirror 401 be successively set on the first imaging optical path, wherein:
The side of the first lens 403 is provided with Electrocontrolled sample platform 104, and opposite side is provided with the second lens 402;
Second lens 402 arrange automatically controlled tilting mirror 401 away from the side of this Electrocontrolled sample platform 104.
In the embodiment of the present invention, by adjusting the position of the first lens 403 and the second lens 402, automatically controlled tilting mirror 401 is made to become conjugate relation with the entrance pupil of image-forming objective lens 105.The automatically controlled tilting mirror 401 of timely adjustment makes imaging shooting area Sp movement fast on moving object sample, to remain overlap to make imaging shooting area Sp with plan shooting area Si, thus the motion eliminating moving object sample is on the impact of super-resolution imaging.Also can change the position of intending shooting area Si fast by controlling Electrocontrolled sample platform in the embodiment of the present invention, thus reach the object that imaging shooting area Sp overlaps with plan shooting area Si position.Those skilled in the art can select according to concrete occasion, and the present invention is not construed as limiting.
As a kind of concrete example of super-resolution image-forming module 30, as shown in Figure 2, the super-resolution image-forming module 30 that the embodiment of the present invention provides is connected with position feedback control module 40, by adjusting the angle of automatically controlled tilting mirror 401 to adjust the imaging shooting area Sp that the first imaging optical path changes super-resolution image-forming module 30; , be also connected with computing equipment 202 meanwhile, receive the imaging frame frequency f transmitted from computing equipment 202
s, according to this imaging frame frequency f
ssuper-resolution imaging is carried out to plan imaging shooting area Si, thus the picture rich in detail in moving object sample moving process can be taken.
In practical application, super-resolution image-forming module 30 can from the equipment such as adopting photoactivation to locate microtechnic (PALM), random optical reconstruct microtechnic (STORM) or Structured Illumination microtechnic (SIM), also the high-resolution imaging equipment such as the focusing of laser scanning light, two (many) photon fluorescence micro-imaging can be adopted, those skilled in the art can select according to concrete use scenes, and the present invention is not construed as limiting.
That introduces that the embodiment of the present invention provides below is a kind of for the super-resolution formation method of fast moving objects and the course of work of device.
As shown in Figure 3, moving object sample A is placed in the sample cell in sample stage 104.
The light that light fixture 101 sends is incident upon bottom sample cell after the 3rd lens 102 and condenser 103.The image that moving object sample A and super-resolution imageable target B1, B2 and B3 are formed after image-forming objective lens 105 and optical splitter 106, then is transferred to rapid image collecting device 201 after imaging len 107 and optical filtering 108.
These rapid image collecting device 201 Quick Acquisition at least two images are also transferred to computing equipment 202, arranged the region of interest ROI comprising super-resolution imageable target B1, B2 and B3 according to the position of super-resolution imageable target B1, B2 and B3 on moving object sample by this computing equipment 202, rower of going forward side by side is fixed.
This computing equipment 202 obtains the position of moving object sample at least two images and super-resolution imageable target; Contrast the position of moving object sample at least two images and super-resolution imageable target respectively, to obtain the displacement of moving object sample and super-resolution imageable target; According to the displacement of moving object sample and super-resolution imageable target and imaging frame frequency f when obtaining at least two images
sobtain the movement velocity V of moving object sample A, the movement velocity V of region of interest ROI
rand the movement velocity V of super-resolution imageable target
i.Wherein, imaging frame frequency f
sobtain the movement velocity V of moving object sample A, intend the movement velocity V of shooting area Si
r, super-resolution imageable target movement velocity V
iwith the resolution PPI of super-resolution image-forming module
sthe following condition of demand fulfillment:
|V
r-V
i|/f
s≤PPI
s
Computing equipment 202 obtains imaging rate according to above-mentioned formula and is transferred to super-resolution image-forming module 30, and according to obtaining the movement velocity V of moving object sample A, intending the movement velocity V of shooting area Si
r, super-resolution imageable target movement velocity V
ithe angle obtaining automatically controlled tilting mirror 401 is transferred to position feedback control module 40.
Position feedback control module 40 adjusts the angle of automatically controlled tilting mirror 401 according to received angle information, make imaging shooting area Sp movement fast on moving object sample to adjust automatically controlled tilting mirror 401 in time, to remain with plan shooting area Si to make imaging shooting area Sp and overlap.
By positional information being transferred to the position that Electrocontrolled sample platform 104 adjusts sample, imaging shooting area Sp can also being made to keep overlapping with plan shooting area Si, thus eliminates the movement velocity V intending shooting area Si in the embodiment of the present invention
rimpact.
The imaging rate of high-resolution imaging module 30 is usually relevant to the size of its imaging shooting area Sp; When resolution is constant, Sp is larger for imaging shooting area, imaging frame frequency f
sless.Super-resolution image-forming module 30 is according to the resolution PPI of super-resolution image-forming module
s, intend the movement velocity V of shooting area Si
rand the movement velocity V of super-resolution imageable target
iobtain the imaging frame frequency f of super-resolution image-forming module
sand imaging shooting area Sp, and then reduce or eliminate (V
i-V
r) impact.Then high-resolution imaging module 30 carries out super-resolution imaging according to the size of imaging shooting area Sp to super-resolution imageable target on moving object sample A.
In sum, the super-resolution formation method for fast moving objects that the embodiment of the present invention provides and device, by obtaining position and the movement velocity of moving object sample and super-resolution imageable target, and the area of imaging shooting area is obtained in conjunction with the resolution of super-resolution image-forming module, ensure super-resolution imaging rate; Keep overlapping with the position made with sample is intended shooting area by the position be adjusted to as shooting area, thus the motion of elimination moving object sample is on the impact of super-resolution imaging.The present invention can carry out super-resolution imaging to moving object quickly and automatically, also can the morphosis of automatic analysis moving object, is particularly useful for the rapid ultra high resolution imaging of the various Moving Objects such as spermatoblast, biological tissue.
Also it should be noted that, in this article, the such as relational terms of first and second grades and so on is only used for an entity or operation to separate with another entity or operational zone, and not necessarily requires or imply the relation that there is any this reality between these entities or operation or sequentially.And, term " comprises ", " comprising " or its any other variant are intended to contain comprising of nonexcludability, thus make to comprise the process of a series of key element, method, article or equipment and not only comprise those key elements, but also comprise other key elements clearly do not listed, or also comprise by the intrinsic key element of this process, method, article or equipment.
Although describe embodiments of the present invention by reference to the accompanying drawings, but those skilled in the art can make various modifications and variations without departing from the spirit and scope of the present invention, such amendment and modification all fall into by within claims limited range.
Claims (6)
1., for a super-resolution formation method for fast moving objects, it is characterized in that, comprising:
Position according to super-resolution imageable target on moving object sample arranges the region of interest ROI comprising described super-resolution imageable target;
Obtain the movement velocity V of described super-resolution imageable target
iwith the movement velocity V of described region of interest ROI
rwith position;
According to the resolution PPI of super-resolution image-forming module
s, described region of interest ROI movement velocity V
rand the movement velocity V of described super-resolution imageable target
iobtain the imaging frame frequency f of described super-resolution image-forming module
s;
According to described imaging frame frequency f
sthe size of described imaging shooting area Sp is calculated with the relation of imaging shooting area Sp; Sample defines the plan shooting area Si that comprises super-resolution imaging target, the size of described plan shooting area Si and the measure-alike of described imaging shooting area Sp, and with region of interest ROI, there is identical movement velocity V
r;
Adjust the position of described plan shooting area Si or described imaging shooting area Sp, keep overlapping with described imaging shooting area Sp to make described plan shooting area Si; And according to described plan shooting area Si, super-resolution imaging is carried out to super-resolution imageable target on moving object sample.
2. super-resolution formation method according to claim 1, is characterized in that, described in
According to the resolution PPI of super-resolution image-forming module
s, described region of interest ROI movement velocity V
rand the movement velocity V of described super-resolution imageable target
iobtain the imaging frame frequency f of described super-resolution image-forming module
sstep in adopt following formula to obtain described imaging frame frequency f
s:
|V
r-V
i|/PPI
s≤f
s。
3. super-resolution formation method according to claim 1, is characterized in that, the movement velocity V of the described super-resolution imageable target of described acquisition
iwith the movement velocity V of region of interest ROI
rwith position by obtaining with under type:
At least two images of region of interest ROI are gathered respectively in Preset Time;
The position of moving object sample and super-resolution imageable target at least two images described in contrast, to obtain the displacement of moving object sample and super-resolution imageable target;
The movement velocity V of described region of interest ROI is obtained according to the displacement of moving object sample and super-resolution imageable target and imaging frame frequency
rand the movement velocity V of described super-resolution imageable target
i.
4. super-resolution formation method according to claim 1, is characterized in that, the movement velocity V of the described super-resolution imageable target of described acquisition
iwith the movement velocity V of region of interest ROI
robtain in the following manner with position:
In region of interest ROI selected one with first representative point that can represent its motion feature, super-resolution imageable target is selected the second representative point that can represent its motion feature;
The light intensity of described first representative point and described second representative point is gathered respectively in the different moment;
The variation relation of the light intensity time of the first representative point described in comparative analysis and described second representative point, to obtain displacement and the speed of moving object sample area-of-interest and super-resolution imageable target.
5. super-resolution formation method according to claim 1, it is characterized in that, the position of described adjustment described plan shooting area Si or described imaging shooting area Sp, to make described plan shooting area Si keep in the step overlapped with described imaging shooting area Sp, realize described plan shooting area Si in the following manner and keep overlapping with described imaging shooting area Sp:
According to the movement velocity V of described region of interest ROI
radjust the position of described imaging shooting area Sp, make that the motion of described imaging shooting area Sp is identical with described region of interest ROI direction of motion, velocity magnitude is equal.
6. super-resolution formation method according to claim 1, it is characterized in that, the position of described adjustment described plan shooting area Si or described imaging shooting area Sp, to make described plan shooting area Si keep in the step overlapped with described imaging shooting area Sp, realize described plan shooting area Si in the following manner and keep overlapping with described imaging shooting area Sp:
Described imaging shooting area Sp remains unchanged, according to the movement velocity V of described region of interest ROI
radjustment sample cell, produces the movement velocity-V contrary with described region of interest ROI direction of motion, velocity magnitude is equal to make sample
r.
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