CN105319725B - Super-resolution imaging method for fast moving objects - Google Patents
Super-resolution imaging method for fast moving objects Download PDFInfo
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- CN105319725B CN105319725B CN201510728880.9A CN201510728880A CN105319725B CN 105319725 B CN105319725 B CN 105319725B CN 201510728880 A CN201510728880 A CN 201510728880A CN 105319725 B CN105319725 B CN 105319725B
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- 238000003384 imaging method Methods 0.000 title claims abstract description 141
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- G—PHYSICS
- G02—OPTICS
- G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
- G02B27/00—Optical systems or apparatus not provided for by any of the groups G02B1/00 - G02B26/00, G02B30/00
- G02B27/58—Optics for apodization or superresolution; Optical synthetic aperture systems
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- G—PHYSICS
- G06—COMPUTING; CALCULATING OR COUNTING
- G06V—IMAGE OR VIDEO RECOGNITION OR UNDERSTANDING
- G06V10/00—Arrangements for image or video recognition or understanding
- G06V10/20—Image preprocessing
- G06V10/25—Determination of region of interest [ROI] or a volume of interest [VOI]
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- G—PHYSICS
- G06—COMPUTING; CALCULATING OR COUNTING
- G06T—IMAGE DATA PROCESSING OR GENERATION, IN GENERAL
- G06T2207/00—Indexing scheme for image analysis or image enhancement
- G06T2207/10—Image acquisition modality
- G06T2207/10056—Microscopic image
Abstract
The present invention relates to a kind of super-resolution imaging method for fast moving objects.This method includes：The region of interest ROI for including super-resolution imageable target is set according to the position of super-resolution imageable target on moving object sample；The movement velocity of super-resolution imageable target and ROI movement velocity and position are obtained, and obtains the imaging frame frequency of super-resolution image-forming module；Sp size is calculated according to the relation of imaging frame frequency and imaging shooting area Sp；It is identical with Sp size that a size comprising super-resolution imaging target plan shooting area Si, Si is defined on sample, and there is identical movement velocity with ROI；Shooting area Si is intended according to ROI motion adjustment or is imaged shooting area Sp position, keeps overlapping with imaging shooting area Sp so as to intend shooting area Si；And super-resolution imaging is carried out to super-resolution imageable target on moving object sample according to shooting area Si is intended.The present invention can eliminate the influence that object of which movement is imaged to super-resolution.
Description
Technical field
The present invention relates to acquisition technology field, more particularly to a kind of super-resolution for fast moving objects to be imaged
Method.
Background technology
Generally, the size of the discernmible smallest object of human eye is about 0.1mm.If want to see smaller thing
Body, then need by means of microtechnic.1873, German microtechnic expert Ernest ＆ Whitney Abbe disclose light microscope by
In the diffraction effect and finite aperture resolution ratio of light the principle of the limit be present, be exactly the principle " constraint " conventional optical microscope
In the utilization of Nanometer World.
When only having single fluorescence molecule under the microscopical object lens visual field, it is fitted by specific algorithm, it is easy to more than light
Learn resolution limit.To explore microcosmos, the super-resolution microtechnic for breaking through the optical limit of light microscope meet the tendency of and
It is raw.Unimolecule tracking technique is incorporated into life science by Barak and Webb first within 1981.Although monomolecular registration
Nanoscale can be reached, but it can not improve resolution ratio of the light microscope when differentiating two or more spot light.
Patterson and Lippincott-Schwartz in 2002 utilizes the mutation of green fluorescent protein (GFP) first
(PA-GFP) movement locus of specified protein in the cell is observed.German Eric Bezig are observantly recognized：Using single point
Sub- Imaging-PAM, with reference to the characteristics of luminescence of this fluorescin, can break through optical resolution the limit --- photoactivation is fixed
Position microtechnic (PALM) is born.PALM imaging method can only be used for observing heterogenous expression albumen, to cellular endogenous albumen
It is helpless.2006, U.S. Howard-Hughes research institute scientist foreign citizen of Chinese origin Zhuan Xiao common vetch experimental group was found：Different wavelength
Switchings of the chemiluminescence molecule Cy5 between fluorescence excitation state and dark-state can be controlled.Given this random optical reconstruct is developed
Microtechnic (STORM).Either PALM or STORM super-resolutions microscopy, its point spread function imaging still with
Traditional micro-imaging is consistent, it is necessary to activation-quenching fluorescence molecule repeatedly, so experiment is completed on fixed cells mostly.
2000, Germany scientist Stefan Hell proposed to reduce the spot size of exciting light by physical process, directly
Connect and reduce the halfwidth of point spread function and improve resolution ratio, successfully have developed stimulated emission depletion microtechnic (STED).Change
Become point spread function and realize that another method for breaking through optical diffraction limit is saturated structures illumination microtechnic (SSIM).2005
Year, nonlinear organization illumination optical part is incorporated on traditional microscope by Gustafsson first, has obtained resolution ratio
Reach 50nm image.But existing super-resolution techniques image taking speed is slow, and it is difficult to shooting motion (particularly quick fortune
It is dynamic) sample.
The content of the invention
The present invention one of purpose be to provide a kind of super-resolution imaging method for fast moving objects, with
Solves the technical problem that image taking speed in the prior art is difficult to shoot moving object, particularly biological living slowly.
For achieving the above object, the embodiments of the invention provide a kind of super-resolution for fast moving objects into
Image space method, including：
Set according to the position of super-resolution imageable target on moving object sample and include the super-resolution imageable target
Region of interest ROI；
Obtain the movement velocity V of the super-resolution imageable targetiWith the movement velocity V of the region of interest ROIrWith
Position；
According to the resolution ratio PPI of super-resolution image-forming modules, the region of interest ROI movement velocity VrIt is and described
The movement velocity V of super-resolution imageable targetiObtain the imaging frame frequency f of the super-resolution image-forming modules；
According to the imaging frame frequency fsRelation with being imaged shooting area Sp calculates the size of the imaging shooting area Sp；
Define a plan shooting area Si comprising super-resolution imaging target on sample, the size for intending shooting area Si with it is described into
As shooting area Sp size is identical, and there is identical movement velocity V with region of interest ROIr；
The plan shooting area Si or the imaging shooting area Sp position are adjusted, so that the plan shooting area Si
Keep overlapping with the imaging shooting area Sp；And according to the plan shooting area Si to super-resolution on moving object sample into
As target carries out super-resolution imaging.
Alternatively, it is described
According to the resolution ratio PPI of super-resolution image-forming modules, the region of interest ROI movement velocity VrIt is and described
The movement velocity V of super-resolution imageable targetiObtain the imaging frame frequency f of the super-resolution image-forming modulesThe step of in use with
Lower formula obtains the imaging frame frequency fs：
|Vr-Vi|/PPIs≤fs。
Alternatively, the movement velocity V for obtaining the super-resolution imageable targetiWith the motion of region of interest ROI
Speed VrIt can be obtained in the following manner with position：
Gather at least two images of region of interest ROI respectively in preset time；
The position of moving object sample and super-resolution imageable target at least two images described in contrast, to obtain motion
The displacement of object sample and super-resolution imageable target；
The region of interest is obtained according to the displacement of moving object sample and super-resolution imageable target and imaging frame frequency
Domain ROI movement velocity VrAnd the movement velocity V of the super-resolution imageable targeti。
Alternatively, the movement velocity V for obtaining the super-resolution imageable targetiWith the motion of region of interest ROI
Speed VrObtained in the following manner with position：
One is selected in region of interest ROI to represent a little with the first of its motion feature can be represented, in super-resolution
The second of its motion feature can be represented and represent a little by selecting one in imageable target；
Described first is gathered different at the time of respectively represent point and represent the light intensity put with described second；
First represents point the light intensity of point is represented with described second and change with time relation described in comparative analysis, is transported with obtaining
The displacement of animal body samples area-of-interest and super-resolution imageable target and speed.
Alternatively, the adjustment plan shooting area Si or the imaging shooting area Sp position, so that described
Intend in the step of shooting area Si keeps overlapping with the imaging shooting area Sp, the plan shooting area is accomplished by the following way
Domain Si keeps overlapping with the imaging shooting area Sp：
According to the movement velocity V of the region of interest ROIrThe position of the imaging shooting area Sp is adjusted, is made described
The motion that is imaged shooting area Sp is identical with the region of interest ROI direction of motion, velocity magnitude is equal.
Alternatively, the adjustment plan shooting area Si or the imaging shooting area Sp position, so that described
Intend in the step of shooting area Si keeps overlapping with the imaging shooting area Sp, the plan shooting area is accomplished by the following way
Domain Si keeps overlapping with the imaging shooting area Sp：
The imaging shooting area Sp keeps constant, according to the movement velocity V of the region of interest ROIrAdjust sample
Pond, so that sample produces the movement velocity V with the region of interest ROIr- the V that the direction of motion is opposite, velocity magnitude is equalr。
The embodiment of the present invention is by obtaining the position of moving object sample area-of-interest and super-resolution imageable target
Super-resolution imaging speed is determined with the resolution ratio of movement velocity, and combination super-resolution image-forming module, takes this as a foundation and is adjusted to
As the area of shooting area, ensure super-resolution imaging rate, so as to eliminate motion of the photographic subjects relative to ROI for oversubscription
Distinguish the influence of shooting；By being adjusted to the position as intending shooting area on the position of shooting area or sample, so that imaging is shot
Region keeps overlapping with intending the position of shooting area on sample, so as to eliminate moving object sample ROI motion to super-resolution
The influence of imaging.The present invention quickly and automatically can carry out super-resolution imaging to fast moving objects, can also automatically analyze
The rapid ultra highs of the various Moving Objects such as the morphosis of moving object, the spermatoblast that is particularly suitable for use in, biological tissue differentiate into
Picture.
Brief description of the drawings
The features and advantages of the present invention can be more clearly understood by reference to accompanying drawing, accompanying drawing is schematically without that should manage
Solve to carry out any restrictions to the present invention, in the accompanying drawings：
Fig. 1 is a kind of flow for super-resolution imaging method for fast moving objects that one embodiment of the invention provides
Schematic diagram；
Fig. 2 is that a kind of super-resolution image device structure for fast moving objects that one embodiment of the invention provides shows
It is intended to；
Fig. 3 is to be adjusted to overlap process schematic as shooting area adjusts with plan shooting area in one embodiment of the invention.
Embodiment
With reference to the accompanying drawings and examples, the embodiment of the present invention is described in further detail.Implement below
Example is used to illustrate the present invention, but is not limited to the scope of the present invention.
In a first aspect, the embodiments of the invention provide a kind of super-resolution imaging method for fast moving objects, such as
Shown in Fig. 1, including：
Set according to the position of super-resolution imageable target on moving object sample and include the super-resolution imageable target
Region of interest ROI；
Obtain the movement velocity V of the super-resolution imageable targetiWith the movement velocity V of the region of interest ROIrWith
Position；
According to the resolution ratio PPI of super-resolution image-forming modules, the region of interest ROI movement velocity VrIt is and described
The movement velocity V of super-resolution imageable targetiObtain the imaging frame frequency f of the super-resolution image-forming modules；
According to the imaging frame frequency fsRelation with being imaged shooting area Sp calculates the size of the imaging shooting area Sp；
A plan shooting area Si comprising super-resolution imaging target is defined, the size for intending shooting area Si shoots area with the imaging
Domain Sp size is identical, and has identical movement velocity Vr；
The plan shooting area Si or the imaging shooting area Sp position are adjusted, so that the plan shooting area Si
Keep overlapping with the imaging shooting area Sp；And according to the plan shooting area Si to super-resolution on moving object sample into
As target carries out super-resolution imaging.
The embodiment of the present invention by obtaining position and the movement velocity of moving object sample and super-resolution imageable target,
And the resolution ratio for combining super-resolution image-forming module obtains the area of imaging shooting area, ensures super-resolution imaging rate；It is logical
The position being adjusted to as shooting area or sample is crossed so as to keep overlapping with the position of imaging shooting area plan shooting area, from
And the influence that the motion for eliminating moving object sample is imaged to super-resolution.The present invention can be quickly and automatically to moving object
Super-resolution imaging is carried out, the morphosis of moving object, the spermatoblast that is particularly suitable for use in, biological tissue can also be automatically analyzed
Etc. the rapid ultra high resolution imaging of various Moving Objects.
Alternatively, according to the resolution ratio PPI of super-resolution image-forming modules, the region of interest ROI movement velocity Vr
And the movement velocity V of the super-resolution imageable targetiObtain the imaging frame frequency f of the super-resolution image-forming modulesThe step of
It is middle that imaging frame frequency f is obtained using below equations：
|Vr-Vi|/PPIs≤fs。
Alternatively, the movement velocity V of the super-resolution imageable target is obtainediWith the movement velocity V of region of interest ROIr
Obtained in the following manner with position：
Obtain the position of the moving object sample and super-resolution imageable target at least two images；
Gather at least two images of region of interest ROI respectively in preset time；
The position of moving object sample and super-resolution imageable target at least two images is contrasted, to obtain moving object
The displacement of sample and super-resolution imageable target；
The region of interest is obtained according to the displacement of moving object sample and super-resolution imageable target and imaging frame frequency
Domain ROI movement velocity VrAnd the movement velocity V of the super-resolution imageable targeti。
Alternatively, the movement velocity V of region of interest ROI is obtainedrObtained in the following manner with position：
One is selected in region of interest ROI to represent a little with the first of its motion feature can be represented, in super-resolution
The second of its motion feature can be represented and represent a little by selecting one in imageable target；
The first representative point and second is gathered different at the time of respectively and represents the light intensity of point；
Relation that the representative of comparative analysis first is put and the second light intensity for representing point changes with time, to obtain moving object sample
The displacement of product region of interest ROI and super-resolution imageable target and speed.
Refer to it will be appreciated that above " its motion feature can be represented ", first represents point and region of interest ROI
The direction of motion is identical, velocity magnitude is equal.In practical application, movement velocity and the position of the point are obtained by position sensor.
It is real in the following manner to make imaging shooting area Sp keep overlapping with region of interest ROI in practical application
It is existing：
According to the movement velocity V of region of interest ROIrIt is adjusted to, as shooting area Sp motion feature, shoot imaging
Region Sp is identical with the region of interest ROI direction of motion, velocity magnitude is equal.Or imaging shooting area Sp keeps constant, root
According to the movement velocity V of region of interest ROIrSample cell is adjusted, so that sample produces and the region of interest ROI direction of motion
Conversely, the equal movement velocity-V of velocity magnituder。
To embody a kind of the superior of super-resolution imaging method for fast moving objects provided in an embodiment of the present invention
Property, the embodiment of the present invention additionally provides a kind of super-resolution imaging side device for fast moving objects, as shown in Fig. 2 bag
Include：
Conventional resolution micro-imaging module 10, for showing moving object sample and super-resolution imageable target to provide
The position and movable information of the moving object sample and super-resolution imageable target；
Rapid image gathers and processing module 20, for obtaining movement velocity V, the super-resolution imaging of moving object sample
The movement velocity V of targeti, intend shooting area Si movement velocity VrWith position and imaging shooting area Sp size, and provide
To position feedback control module 40 and super-resolution image-forming module 30；
Position feedback control module 40, for adjusting the imaging shooting area Sp or intending imaging region Si position,
Keep overlapping to intend shooting area Si with being imaged shooting area Sp；
Super-resolution image-forming module 30, for according to super-resolution imaging rate requirement, being adjusted to as shooting area Sp chi
It is very little, super-resolution imaging is carried out to super-resolution imageable target on moving object sample.
As a kind of specific example of conventional resolution micro-imaging module 10, as shown in Fig. 2 the embodiment of the present invention provides
Conventional resolution micro-imaging module 10 include lighting unit and imaging unit.Lighting unit includes being successively set on the second one-tenth
As the lighting apparatus 101 in light path, the 3rd lens 102 and condenser 103, wherein, the side of the 3rd lens 102 is provided with illumination
Equipment 101, opposite side are provided with condenser 103；It is provided with the side of the remote lighting apparatus 101 of the condenser 103 automatically controlled
Sample stage 104.Imaging unit includes being successively set on image-forming objective lens 105 on the second imaging optical path, optical splitter 106 and the three one-tenth
As the imaging len 107 in light path, optical filtering 108；The side of image-forming objective lens 105 is provided with Electrocontrolled sample platform 104, and opposite side is set
It is equipped 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 set
Put in the side of optical splitter 106, opposite side is provided with optical filtering 108.
In practical application, those skilled in the art according to specific usage scenario, can select the routine point of suitable parameters
Resolution micro-imaging module shows that fast moving objects sample and super-resolution imageable target, the present invention are not construed as limiting to realize.
As a kind of collection of rapid image and the specific example of processing module 20, as shown in Fig. 2 the embodiment of the present invention provides
Rapid image collection with processing module 20 include rapid image collecting device 201 and computing device 202, wherein：
Rapid image collecting device 201, for gathering moving object sample and super-resolution imageable target to provide the fortune
The position and movable information of animal body samples and super-resolution imageable target；
Computing device 202, for the position and movable information according to moving object sample and super-resolution imageable target with
Obtain movement velocity V, the movement velocity V of super-resolution imageable target of moving object samplei, intend shooting area Si motion speed
Spend VrWith position and imaging shooting area Sp size.
In practical application, rapid image collecting device 201 can use such as charge coupled cell (Charge-coupled
Device, CCD) or complementary metal oxide semiconductor (Complementary Metal Oxide Semiconductor,
The imaging sensor such as CMOS), also can be used Position-Sensitive Detector etc., and those skilled in the art can be according to specifically using field
Scape is selected, and the present invention is not construed as limiting.
In practical application, ambient noise of the computing device 202 at least two images, moving object sample A forms
Similitude of structure, moving object sample A and background etc., take the technologies such as different background modelings, target identification, Track Pick-up
Movement velocity is calculated, those skilled in the art can select suitable processing method according to specific usage scenario.
As a kind of specific example of position feedback control module 40, as shown in Fig. 2 position provided in an embodiment of the present invention
Feedback control module 40 includes the first lens 403, the second lens 402 and the automatically controlled tilting mirror being successively set on the first imaging optical path
401, wherein：
The side of first lens 403 is provided with Electrocontrolled sample platform 104, and opposite side is provided with the second lens 402；
Side of second lens 402 away from the Electrocontrolled sample platform 104 sets automatically controlled tilting mirror 401.
In the embodiment of the present invention, by adjusting the position of the first lens 403 and the second lens 402, make automatically controlled tilting mirror 401 and
The entrance pupil of image-forming objective lens 105 is into conjugate relation.Adjusting automatically controlled tilting mirror 401 in time makes imaging shooting area Sp in moving object sample
It is upper quick mobile, overlapped so that imaging shooting area Sp remains with plan shooting area Si, so as to eliminate moving object sample
The influence that is imaged to super-resolution of motion.It can also be clapped in the embodiment of the present invention by controlling Electrocontrolled sample platform quickly to change plan
Region Si position is taken the photograph, the purpose overlapped so as to reach imaging shooting area Sp with intending shooting area Si positions.Art technology
Personnel can be selected according to specific occasion, and the present invention is not construed as limiting.
As a kind of specific example of super-resolution image-forming module 30, as shown in Fig. 2 superelevation provided in an embodiment of the present invention
Resolution imaging module 30 is connected with position feedback control module 40, and first is adjusted by adjusting the angle of automatically controlled tilting mirror 401
Imaging optical path changes the imaging shooting area Sp of super-resolution image-forming module 30；Meanwhile be also connected with computing device 202, receive
The imaging frame frequency f transmitted from computing device 202s, according to imaging frame frequency fsSuperelevation point is carried out to intending imaging shooting area Si
Imaging is distinguished, so as to shoot the picture rich in detail in moving object sample moving process.
In practical application, super-resolution image-forming module 30 can from for example with photoactivation position microtechnic (PALM),
The equipment of random optical reconstruct microtechnic (STORM) or Structured Illumination microtechnic (SIM), can also use laser scanning
The high-resolution imaging equipment such as light focusing, double (more) photon fluorescence micro-imagings, those skilled in the art can be according to specifically making
Selected with scene, the present invention is not construed as limiting.
A kind of super-resolution imaging method and dress for fast moving objects provided in an embodiment of the present invention is described below
The course of work put.
As shown in figure 3, moving object sample A is placed in the sample cell in sample stage 104.
The light that lighting apparatus 101 is sent is incident upon sample cell bottom 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 is by image-forming objective lens 105 and light splitting
After device 106, then rapid image collecting device 201 is transferred to after imaging len 107 and optical filtering 108.
The Quick Acquisition of rapid image collecting device 201 at least two images are simultaneously transferred to computing device 202, by the calculating
Equipment 202 sets according to the position of super-resolution imageable target B1, B2 and B3 on moving object sample and is imaged comprising super-resolution
Target B1, B2 and B3 region of interest ROI, and demarcated.
The computing device 202 obtains the position of the moving object sample and super-resolution imageable target at least two images
Put；The position of the moving object sample and super-resolution imageable target at least two images is contrasted respectively, to obtain moving object
The displacement of body sample and super-resolution imageable target；According to the displacement of moving object sample and super-resolution imageable target and obtain
Take imaging frame frequency f during at least two imagessObtain moving object sample A movement velocity V, region of interest ROI motion
Speed VrAnd the movement velocity V of super-resolution imageable targeti.Wherein, imaging frame frequency fsObtain moving object sample A motion
Speed V, the movement velocity V for intending shooting area Sir, super-resolution imageable target movement velocity ViWith super-resolution image-forming module
Resolution ratio PPIsNeed to meet following condition：
|Vr-Vi|/fs≤PPIs
Computing device 202 obtains imaging rate according to above-mentioned formula and is transferred to super-resolution image-forming module 30, and according to
Obtain moving object sample A movement velocity V, intend shooting area Si movement velocity Vr, super-resolution imageable target motion
Speed ViThe angle for 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, with timely
Adjusting automatically controlled tilting mirror 401 makes imaging shooting area Sp quickly be moved on moving object sample so that imaging shooting area Sp with
Intend shooting area Si and remain coincidence.
In the embodiment of the present invention can also by the way that positional information is transferred into the position that Electrocontrolled sample platform 104 adjusts sample,
Imaging shooting area Sp is set to keep overlapping with intending shooting area Si, so as to eliminate the movement velocity V for intending shooting area SirShadow
Ring.
It is related that the imaging rate of high-resolution imaging module 30 is generally imaged shooting area Sp size to it；Differentiating
When rate is constant, imaging shooting area Sp is bigger, imaging frame frequency fsIt is smaller.Super-resolution image-forming module 30 is imaged according to super-resolution
The resolution ratio PPI of modules, intend shooting area Si movement velocity VrAnd the movement velocity V of super-resolution imageable targetiObtain
The imaging frame frequency f of super-resolution image-forming modulesAnd imaging shooting area Sp, and then reduce or eliminate (Vi-Vr) influence.
Then high-resolution imaging module 30 according to imaging shooting area Sp size to super-resolution imageable target on moving object sample A
Carry out super-resolution imaging.
In summary, the super-resolution imaging method and device provided in an embodiment of the present invention for fast moving objects,
By obtaining position and the movement velocity of moving object sample and super-resolution imageable target, and combine super-resolution imaging mould
The resolution ratio of block obtains the area of imaging shooting area, ensures super-resolution imaging rate；By being adjusted to as shooting area
Position with intending the position of shooting area on sample so as to keep overlapping, so as to eliminate the motion of moving object sample to super-resolution
The influence of imaging.The present invention quickly and automatically can carry out super-resolution imaging to moving object, can also automatically analyze motion
The rapid ultra high resolution imaging of the various Moving Objects such as the morphosis of object, the spermatoblast that is particularly suitable for use in, biological tissue.
It should also be noted that, herein, such as first and second or the like relational terms are used merely to one
Entity or operation make a distinction with another entity or operation, and not necessarily require or imply between these entities or operation
Any this actual relation or order be present.Moreover, term " comprising ", "comprising" or its any other variant are intended to contain
Lid nonexcludability includes, so that process, method, article or equipment including a series of elements not only will including those
Element, but also the other element including being not expressly set out, or it is this process, method, article or equipment also to include
Intrinsic key element.
Although being described in conjunction with the accompanying embodiments of the present invention, those skilled in the art can not depart from this hair
Various modifications and variations are made in the case of bright spirit and scope, such modifications and variations are each fallen within by appended claims
Within limited range.
Claims (5)
- A kind of 1. super-resolution imaging method for fast moving objects, it is characterised in that including：The sense for including the super-resolution imageable target is set according to the position of super-resolution imageable target on moving object sample Interest region ROI；Obtain the movement velocity V of the super-resolution imageable targetiWith the movement velocity V of the region of interest ROIrWith position；According to the resolution ratio PPI of super-resolution image-forming modules, the region of interest ROI movement velocity VrAnd the superelevation The movement velocity V of resolution imaging targeti, using the imaging frame frequency f of the below equation acquisition super-resolution image-forming modules:|Vr-Vi|/PPIs≤fs；According to the imaging frame frequency fsRelation with being imaged shooting area Sp calculates the size of the imaging shooting area Sp；In sample A plan shooting area Si comprising super-resolution imaging target is defined on product, the size for intending shooting area Si is clapped with the imaging It is identical to take the photograph region Sp size, and there is identical movement velocity V with region of interest ROIr；The plan shooting area Si or the imaging shooting area Sp position are adjusted, so that the plan shooting area Si and institute Imaging shooting area Sp is stated to keep overlapping；And mesh is imaged to super-resolution on moving object sample according to the plan shooting area Si Mark carries out super-resolution imaging.
- 2. super-resolution imaging method according to claim 1, it is characterised in that described to obtain the super-resolution imaging The movement velocity V of targetiWith the movement velocity V of region of interest ROIrIt can be obtained in the following manner with position：Gather at least two images of region of interest ROI respectively in preset time；The position of moving object sample and super-resolution imageable target at least two images described in contrast, to obtain moving object The displacement of sample and super-resolution imageable target；The region of interest ROI is obtained according to the displacement of moving object sample and super-resolution imageable target and imaging frame frequency Movement velocity VrAnd the movement velocity V of the super-resolution imageable targeti。
- 3. super-resolution imaging method according to claim 1, it is characterised in that described to obtain the super-resolution imaging The movement velocity V of targetiWith the movement velocity V of region of interest ROIrObtained in the following manner with position：One is selected in region of interest ROI and is represented a little with the first of its motion feature can be represented, is imaged in super-resolution The second of its motion feature can be represented and represent a little by selecting one in target；Described first is gathered different at the time of respectively represent point and represent the light intensity put with described second；Relation that first representative described in comparative analysis is put and the described second light intensity for representing point changes with time, to obtain moving object The displacement of body sample area-of-interest and super-resolution imageable target and speed.
- 4. super-resolution imaging method according to claim 1, it is characterised in that the adjustment plan shooting area Si Or the position of the imaging shooting area Sp, so that the plan shooting area Si keeps overlapping with the imaging shooting area Sp The step of in, the plan shooting area Si is accomplished by the following way and keeps overlapping with the imaging shooting area Sp：According to the movement velocity V of the region of interest ROIrThe position of the imaging shooting area Sp is adjusted, claps the imaging The motion of taking the photograph region Sp is identical with the region of interest ROI direction of motion, velocity magnitude is equal.
- 5. super-resolution imaging method according to claim 1, it is characterised in that the adjustment plan shooting area Si Or the position of the imaging shooting area Sp, so that the plan shooting area Si keeps overlapping with the imaging shooting area Sp The step of in, the plan shooting area Si is accomplished by the following way and keeps overlapping with the imaging shooting area Sp：The imaging shooting area Sp keeps constant, according to the movement velocity V of the region of interest ROIrSample cell is adjusted, with Sample is set to produce the movement velocity-V opposite with the region of interest ROI direction of motion, velocity magnitude is equalr。
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