CN106768396A - A kind of method and system based on differential contrast imaging reduction quantitative phase images - Google Patents
A kind of method and system based on differential contrast imaging reduction quantitative phase images Download PDFInfo
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Abstract
The invention discloses a kind of method and system based on differential contrast image Quantitative reduction phase image, it is related to computer generated image field.The present invention establishes the target image of stabilization to the method for reduction quantitative phase images, and the present invention initially sets up differential contrast and is imaged two-dimension optical phase transfer function H (u), and then sets up H (u), the frequency-domain function of differential contrast imageAnd the relation between quantitative phase information, it is final to realize that carrying out de-convolution operation by the phase transfer function being imaged to differential contrast recovers quantitative phase information, further the present invention have studied multiaxis and divide the method to form Quantitative reduction phase information under asymmetric lighting pattern to strengthen the re-configurability of phase information on different directions, and use mathematics to optimize to reduce the increased problem of frequency noise caused by direct deconvolution.Instant invention overcomes the defect that traditional quantitative image acquisition operations are complicated, image-forming condition is harsh, and the image resolution ratio for obtaining is higher.
Description
Technical field
The present invention relates to phase imaging technical field, and in particular to one kind is based on differential contrast image restoring quantitative phase figure
The method of picture and the reduction micro imaging system of the quantitative phase based on LED array light source.
Background technology
In some specific areas, such as optical measurement, materials physics, adaptive optics, X-ray diffraction optics, electronic display
The fields such as micro-, biomedical imaging, most of sample belongs to phase object.The amplitude transmittance distribution of this type objects is equal
It is even, but the spatial distribution of refractive index or thickness is uneven, therefore the light wave-amplitude of phase object changes very small, phase change is non-
Chang great.Human eye or other photo-detectors all can only judgment object amplitude variations and the change of its phase cannot be judged, therefore not
Thickness or the different various pieces of refractive index in phase object can be distinguished.So for these fields, obtaining phase information and seeming
It is particularly important.
Phase contrast (Phase Contrast) microtechnic as phase imaging technology milestone, its general principle be by
The bias light of illumination is separated with sample scattering light, so as to represent foreground detail so that phase place change is with the shape of Strength Changes
Formula is showed.But there is phase reversing, gradually halo and dark effect, thickness of sample, cover glass and load in traditional phase contrast microtechnic
The deficiency of the aspects such as slide influence.To solve these problems, differential interference contrast microscopes is occurred in that in succession on the basis of phase contrast microtechnic
Than being imaged series of new techniques such as (DPC) into (DIC), differential contrast.But these technologies have complex operation, image-forming condition is harsh
Etc. defect, what is more important these methods cannot obtain global quantitative phase information.
Quantitative phase imaging is imaged and detected by the phase delay of measuring beam come the refraction performance to sample.
In optical measurement, because the frequency of light wave is very high, detector cannot direct detection light phase distribution.In practical operation, can
Data measured directly are often the intensity distribution of wave field, therefore, phase recovery is of optical measurement and imaging technique
Important topic, how to obtain phase information from intensity measurement data turns into technical problem to be solved in research.
The content of the invention
In view of mentioned above, the present invention proposes a kind of quantitative phase reduction micro imaging system to solve above-mentioned technical problem
And the method based on differential contrast image restoring quantitative phase images.
Therefore, the present invention is adopted the following technical scheme that：
On the one hand the present invention provides a kind of quantitative phase reduction micro imaging system, including transmission electron microscope, its feature
It is that the transmission electron microscope light source is LED array, and the illumination pattern of the LED array passes through programming Control, also including use
In the multi-dimensional displacement platform of correction LED array light source position, gather the camera installation of microscopic result and image will be gathered
It is reduced to the processing equipment of quantitative phase images.
Further, LED array of the present invention is the rectangular array being equidistantly arranged to make up by m × n light emitting diode, m,
N is positive integer；Lighting for LED array illumination pattern uses static screen pattern.
Further, present invention additionally comprises control device, the controller connects with LED array light source and camera installation respectively
Connect synchronous with camera frequency acquisition using time division multiplexing completion illumination pattern switching to realize；Specifically, the time-division is multiple
Refer to that single control device can be respectively completed signal triggering and the phase that LED array illumination pattern switches in different time sections with pattern
Machine gather signal triggering, can so save controlling switch resource, because the triggered time is extremely short, should not influence pattern switch with
The synchronization of camera collection.Further, processing equipment of the present invention can complete the derivation of view data and the calculating of view data
With treatment, the final quantitative phase imaging picture for obtaining sample；Specifically, the embodiment of the present invention uses what is matched with camera installation
Software completes the triggering of IMAQ and the derivation of image, at the image with same treatment effect such as MATLAB or VC++
Science and engineering tool completes the calculating of view data；
The specific processing method of processing equipment of the present invention is as follows：
Step A：Obtain the differential contrast imaging picture I that sample is divided with any one symmetrical direction of principal axisDPC；Define first
The first object image gathered under difference illumination light is I1, the second target image under the second difference illumination light is I2, then IDPC's
Preparation method is：
Step B：Differential contrast is set up based on weak signal target transmission function and is imaged two-dimension optical phase transfer function H (u), its table
It is as follows up to formula：
H (u)=i [∫ ∫ S (u ') P* (u ') p (u '+u) d2u′-∫∫s(u′)P*(u′)p(u′-u)d2u′]
Wherein, S (u ') is light source intensity distribution function, and P (u ') is pupil transmission function, and u ' is coordinate in pupil plane；
Step C：Set up differential contrast image in Fourier as follows with phase information relational model：
Then the quantitative phase information of reduction is on Fourier：
It is rightRow inverse Fourier transform, is finally obtained quantitative phase information.
Because above-mentioned differential contrast imaging results are, with the differential contrast image of single symmetrical direction of principal axis acquisition, not wrap completely
Containing the phase information on symmetry axis；It is phase reconstruction ability on enhancing different directions, can be along any direction using many
Axle divides to form asymmetric lighting pattern, is imaged picture so as to obtain the differential contrast comprising different directions phase information, then enter
One step completes the differential contrast imaging containing out of phase information and is reduced to quantitative phase picture.
Based on mentioned above, the present invention provides multiple symmetry axis and divides to be formed obtain determining for sample under asymmetric lighting pattern
The processing method of phase imaging picture is measured, it is specific as follows：
Step A：Obtain the differential contrast imaging picture that sample any two or multiple symmetrical direction of principal axis are divided
Definition is divided under illumination pattern with first symmetry axis, and the first object image of light collection is illuminated using the first difference
It is I11, the second target image for using the second difference to illuminate light collection is I12；Definition further divides photograph with j-th strip symmetry axis
Under bright pattern, the first object image for using the first difference to illuminate light collection is Ij1, using the of the second difference illumination light collection
Two target images are Ij2；Then differential contrast image is：
Step B：Differential contrast is set up based on weak signal target transmission function and is imaged two-dimension optical phase transfer function H (u), its table
It is as follows up to formula：
H (u)=i [∫ ∫ s (u ') P*(u′)p(u′+u)d2u′-∫∫S(u′)P*(u′)p(u′-u)d2u′]
Wherein, S (u ') is light source intensity distribution function, and P (u ') is pupil transmission function, and u ' is coordinate in pupil plane；
Step C：Set up differential contrast image in Fourier as follows with phase information relational model：
Aforesaid equation is combined and is solved, then calculated by equation below based on the quantitative phase information that multiaxis is divided：
Due to H (u) and the Frequency point axially along intersection be zero and participate in computing frequency it is wider, therefore directly deconvolution
The noise in these frequencies can be significantly superimposed, it is possible to which, by regularization least square method solve problem, final abbreviation is quantified
Phase information expression formula is as follows：Wherein, α is regularization parameter
On the other hand the present invention provides a kind of method based on differential contrast image restoring quantitative phase images, wherein for
The division of asymmetric lighting pattern is divided according to any direction using single symmetry axis, is comprised the following steps：
Step A：Using the LED array of programming Control as light source, shone based on time division multiplexing control LED array light source
The switching of bright pattern gathers the Frequency Synchronization of image with camera installation, and wherein target image is carried out by asymmetric lighting pattern
Collection, the asymmetric lighting pattern is to divide to form with single symmetry axis；Define first gathered under the first difference illumination light
Target image is I1, the second target image under the second difference illumination light is I2, then differential contrast image be；
Step B：Differential contrast is set up based on weak signal target transmission function and is imaged two-dimension optical phase transfer function H (u), its table
It is as follows up to formula：
H (u)=i [∫ ∫ S (u ') P* (u ') p (u '+u) d2u′-∫∫S(u′)P*(u′)p(u′-u)d2u′]
Wherein, S (u ') is light source intensity distribution function, and P (u ') is pupil transmission function, and u ' is coordinate in pupil plane；
Step C：The model for setting up Fourier's collection of illustrative plates energy of differential contrast image is as follows：
Then the quantitative phase information of reduction is on Fourier：
It is rightInverse Fourier transform is carried out, it is final to obtain quantitative phase information.
Because above-mentioned differential contrast imaging results are, with the differential contrast image of single symmetrical direction of principal axis acquisition, not wrap completely
Containing the phase information on symmetry axis；It is phase reconstruction ability on enhancing different directions, can be along any direction using many
Axle divides to form asymmetric lighting pattern, is imaged picture so as to obtain the differential contrast comprising different directions phase information, then enter
One step completes the differential contrast imaging containing out of phase information and is reduced to quantitative phase picture.
Based on a kind of mentioned above, method based on differential contrast image restoring quantitative phase images of present invention offer, its
In for the division of asymmetric lighting pattern divided according to any direction using two or more symmetry axis, including with
Lower step：
Step A：Using the LED array of programming Control as light source, based on time division multiplexing control LED array light source
Illumination pattern gathers the Frequency Synchronization of image with camera installation, and wherein target image is adopted by asymmetric lighting pattern
Collection, the asymmetric lighting pattern is to divide to form with the symmetry axis of two or more any directions；Collection target image is specific
Step operation in the following order：
A1：Illumination pattern is divided using first symmetry axis, the first object figure gathered under the first difference illumination light is defined
As being I11, the second target image of gained is I under the second difference illumination light of collection12；
A2：Illumination pattern is further divided using j-th strip symmetry axis, the first mesh gathered under the first difference illumination light is defined
Logo image is Ij1, the second target image gathered under the second difference illumination light is Ij2；
A3：The operation of repeat step A2 until reach the target number of axle, then the differential of two width target images of each step gained
Phase contrast image is as follows：
Step B：Differential contrast is set up based on weak signal target transmission function and is imaged two-dimension optical phase transfer function H (u)：
H (u)=i [∫ ∫ S (u ') P*(u′)p(u′+u)d2u′-∫∫S(u′)P*(u′)p(u′-u)d2u′]
Wherein, S (u ') is light source intensity distribution function, and P (u ') is pupil transmission function, and u ' is coordinate in pupil plane；
Step C：Set up differential contrast image in Fourier as follows with phase information relational model：
Aforesaid equation is combined and is solved, then calculated by equation below based on the quantitative phase information that multiaxis is divided：
Due to H (u) and the Frequency point axially along intersection be zero and participate in computing frequency it is wider, therefore directly deconvolution
The noise in these frequencies can be significantly superimposed, it is possible to which, by regularization least square method solve problem, final abbreviation is quantified
Phase information expression formula is as follows：Wherein, α is regularization parameter
The present invention has the advantages that：
System disclosed by the invention can be established based on differential contrast imaging reduction quantitative phase information, and the present invention
Stabilization from target image to the method for Quantitative reduction phase image, overcome that traditional quantitative image acquisition operations are complicated, imaging
The harsh defect of condition, and the image resolution ratio for obtaining is higher；The present invention builds the quantitative phase based on LED array light source also
Former micro imaging system, obtains target image, then using treatment using asymmetric lighting pattern is synchronous with camera installation collection
Equipment carries out calculation process, final to obtain quantitative phase information.The present invention obtains differential contrast and is imaged picture, differential by treatment
Phase contrast imaging picture contains the gradient information of phase, and the present invention initially sets up differential contrast imaging two-dimension optical phase transmission letter
Number H (u) (specific derivation process is as described above), and then set up H (u), the frequency-domain function of differential contrast imageWith it is fixed
Relation between amount phase information, finally realizes carrying out de-convolution operation by the phase transfer function for being imaged differential contrast extensive
Quantitative phase of appearing again information.The present invention is not only limited to divide to form asymmetric lighting pattern using single symmetry axis, further
Ground, the present invention uses and divides to form asymmetric lighting pattern along multiple symmetry axis of any direction, and then obtains multiple containing not
The differential contrast imaging picture of same-phase information, then carries out joint and solves final reduction quantitative phase information to reach enhancing not
The re-configurability of equidirectional upper phase information.Additionally, quantitative phase information formula of the present invention also to being divided based on multiaxis is carried out
Optimization processing.
Brief description of the drawings
Fig. 1 is the image-forming principle schematic diagram of embodiment of the present invention LED array light source micro imaging system；
Fig. 2 is the flow chart that the embodiment of the present invention reduces quantitative phase images using LED array light source micro imaging system；
Fig. 3 is the structural representation that the embodiment of the present invention is based on LED array light source micro imaging system；
Fig. 4 is the light path correcting process figure of embodiment of the present invention LED array light source；
Fig. 5 is embodiment of the present invention phase contrast imaging illumination pattern schematic diagram.
Specific embodiment
Specific embodiment of the invention is further elaborated with reference to description below accompanying drawing, embodiments herein is only used
In the explanation present invention, but it is not limited to the scope of the invention.
It is as shown in Figure 1 the principle schematic of LED array light source micro imaging system, based on the quantitative phase that the present invention is provided
Position reduction micro imaging system, coherent point light sources intensity I (r in the camerac) be：
I(rc)=| ∫ ∫ [∫ ∫ q (r) o (r) exp (- i2 π ru ") d2r]P(u″)exp(-i2πu″·rc)d2u″|2 (1)
Wherein, the grading function of LED array light source is q (r), and the transmission function of testing sample is o (r), remaining optics device
Part transforming function transformation function is P (u "), rcIt is the coordinate of camera focal plane, u is the coordinate of pupil plane；
When the incoherent light source lighting that sample is expanded, then the light distribution on the focal plane of camera should be differential
The non-coherent addition image of spot light, the intensity of the spot light non-coherent addition image of above-mentioned differential is as follows：
I(rc)=∫ ∫ | ∫ ∫ [∫ ∫ q (r) o (r) exp (- i2 π ru ") d2r]P(u″)exp(-i2πu″·rc)d2u″|2d2u′
(2)
By LED array is remote enough as the sample that light source distance microscope is carried in the present invention, therefore, LED array
Each lighting unit is similar to plane wave in the illumination pattern of formation, then the grading function of LED array light source can be represented
Shown in following formula：
Wherein, S (u ') is the intensity distribution function of LED array light source illumination pattern, and u ' is in LED array light source plane
Coordinate；
Sample penetration functionIt is common sense in the field, μ (r) is intensity transmitance, φ
R () is phase change rate, in o (r) ≈ 1- μ (r)+i φ (r), according to weak object approach method linear simplifiation, this is asked approximately equivalent
Topic, ignoring cross term can obtain following expression：
Formula (3) and formula (4) are substituted into formula (2), and both members are carried out into Fourier transformation, obtain light intensity frequency domain
FunctionCan be expressed by following formula：
Contain the information of three aspects in formula (5)：Background, absorbs contrast and phase-contrast：
Wherein, background item B δ (u) is, by all energy sums of imaging system, can be expressed as follows in formula (5)：
B=∫ ∫ S (u ') | P (u ') |2d2u′ (6)
Light intensity delivery function H in formula (5)absU () can be expressed as follows：
Habs(u)=- [∫ ∫ S (u ') P*(u′)p(u′+u)d2u′+∫∫S(u′)P*(u′)p(u′-u)d2u′] (7)
Phase transfer function H in formula (5)phU () can be expressed as follows：
Hph(u)=i [∫ ∫ S (u ') P*(u′)p(u′+u)d2u′-∫∫S(u′)P*(u′)p(u′-u)d2u′] (8)
Based on derivation above, the present invention is closed based on differential contrast image is set up with the of equal value of quantitative phase information
System.In order to derive the specific transmission function of differential contrast imaging, it is necessary to first be calculated using the two images of asymmetric lighting
Differential contrast image；The embodiment of the present invention is illustrated in figure 2 using the quantitative phase reduction micro-imaging based on LED array light source
The flow chart of system reducing quantitative phase images, concrete operations are as follows：
1st, the microscopic system based on LED array light source illumination is built；
2nd, LED array light source position correction；
3rd, LED array light source forms the design of asymmetric lighting pattern；
4th, differential contrast image restoring quantitative phase images；
It is described in detail below in conjunction with Figure of description and embodiment：
Step 1：Build quantitative phase reduction micro imaging system, including transmission electron microscope, the transmission electron microscope
Light source is LED array, and the illumination pattern of the LED array passes through programming Control, also including for correcting LED array light source position
Multi-dimensional displacement platform, gather microscopic result camera installation and by gather image restoring for quantitative phase images place
Reason equipment.
Further, camera is put in microscopic system front end by build process, using camera monotype Zheng Pin≤24Hz's
High speed camera, (being illustrated in figure 3 the structural representation of micro imaging system of the present invention) and connects LED array light using controller
Source and camera, camera acquisition operations is same during so as to the switching of illumination pattern multi-mode and the different mode of realizing LED array light source
Step.
In preferred embodiment, LED array is positioned on multi-dimensional displacement platform, and multi-dimensional displacement platform is placed in microscope load
60~70mm on thing platform.LED array light source any can meet objective lens numerical hole using 32 × 32,16 × 16,8 × 8,4 × 4 etc.
The array sizes of footpath NA values.
Asymmetric lighting pattern of the present invention is semicircle or semi-circular with LED array center as the center of circle or with LED gusts
Arrange centrosymmetric rectangle, illumination pattern is the first illumination pattern and the second illumination pattern, first illumination pattern and described
Central Symmetry of second illumination pattern on LED array, and the first illumination pattern and the second illumination pattern light simultaneously to be formed with
LED array center is circular, the annular or rectangle at midpoint.
It is below that semicircular implementation method with LED array center as the center of circle further illustrates LED gusts with illumination pattern
The relation of the numerical aperture of row selection of dimension and object lens：
The numerical aperture expression formula of micro objective is as follows：
Wherein, n is medium refraction index, n=1 in instantiation in the present invention；θ is the maximum angle into object lens light；R
The maximum radius of micro objective can be entered for the light emitted line of LED array light source, D is LED array light source and micro objective
Spacing.
From below equation, LED array light source forms the least radius R such as following formulas of illumination pattern：
If setting from 10 × and numerical aperture being 0.25 micro objective, actually it is calculated as follows：
Distance is 4mm between the LED array light source any two adjacent LED center that the embodiment of the present invention is used,
Therefore, the array sizes that should be chosen are at least 8 × 8 standard.
After determining light emitting diode matrix light field range size according to selected object lens, to realize that precisely imaging is also needed to LED
The position correction of array light source.
Step 2：The position correction of LED array light source；This step is mainly included LED array light source center and object lens center
Alignment, the horizontality calibration of LED array light source and the deflection calibration of LED array light source；It is illustrated in figure 4 LED array light source
The basic operation of position correction process：
Step 10：Suitable eyepiece and Aviar need to be selected, due to needing in LED array center and object lens phase loop
The heart is aligned, it is therefore desirable to which then selected phase type object lens are rotated to B mouthfuls, this reality microscopical eyepiece as micro objective
Example is applied using Bert human relations lens, Fourier plane i.e. LED array face so can be just seen；
Step 11：Light apart from LED array center；This implementation is lighted with LED array light source center as the center of circle, and radius is two
The illumination pattern of individual light emitting diode is aligned in order to center；
Step 12：By adjusting multi-dimensional displacement platform, LED array center is aligned with object lens phase loop center；
Because the acquisition of differential contrast image is higher with the precision of directivity requirement to LED array light source horizontality, naked eyes
Calibration does not reach the accuracy of requirement, preferably, completes to calibrate using machine vision technique：
Step 21：Camera is built in microscopical side ports；
Step 22：Suitable lens are placed between camera and microscope side ports, and adjust itself and camera relative position,
Can imaging Fourier plane (and LED array face) according to lens Fourier's variation characteristic camera；
Step 23：LED array horizontality and deflection are measured by PC ends camera software, by adjusting multi-dimensional displacement platform
Horizontality is completed with calibration.
After completing the position correction of LED array light source, to realize that differential contrast imaging also needs further to draw light field region
Point.
Step 3：Design LED array light source forms asymmetric lighting pattern；
In preferred embodiment, LED array light source is formed lighting for illumination pattern and is shielded rather than scan mode using static, because
Static state screen lighting mode can increase thang-kng amount.According to differential contrast imaging mechanism, need to use non-right to obtain phase-contrast figure
Illumination pattern is called common sense in the field.In preferred embodiment, the semicircle illumination figure with LED array center as the center of circle is generally used
Sample, additionally, asymmetric lighting pattern of the present invention can also be semi-circular with LED array center as the center of circle or with LED gusts
Arrange centrosymmetric rectangle.
Differential contrast imaging is obtained by division direction of single symmetry axis, illumination pattern is that the first illumination pattern and second are shone
Bright pattern, the first illumination figure and the second illumination pattern are complementary, as：First illumination pattern 1 and second illumination pattern
Central Symmetry on LED array, and the first illumination pattern and the second illumination pattern light to be formed and be with LED array center simultaneously
Circular, the annular or rectangle at midpoint.
As shown in figure 5, this implementation illumination pattern is the semicircle with LED array center as the center of circle, by with paper level
Angular separation is divided into region 1 and region 2 for the single symmetry axis of α, and division direction is not limited to shown in Figure of description, α
Span be 0 °~180 °；As shown in figure 5, obtain differential contrast imaging results with the single symmetry axis division of any angle being
Phase contrast imaging result on+90 ° of directions of sample α.To obtain the phase contrast imaging result in all directions, take and converted along angle [alpha]
Dividing mode, you can obtain along the phase contrast imaging result on+90 ° of directions of α.
Step 4：The treatment of differential contrast image restoring quantitative phase images；
Switching and the Frequency Synchronization of camera exposure time using controller control LED array light source illumination pattern, complete
In the case where single symmetry axis is divided and to form asymmetric pattern illumination, the collection of target image, gathers under the first difference illumination light of definition
First object image is I1, the second target image under the second difference illumination light is I2,；Then computing is carried out using processing equipment
Treatment, specific processing method is as follows：
Step A：Obtain the differential contrast imaging picture I that sample is divided with any one symmetrical direction of principal axisDPC；Define first
The first object image gathered under difference illumination light is I1, the second target image under the second difference illumination light is I2, then IDPC's
Preparation method is：
For an aberrationless system, pupil function is that a radius is set to target value, the circular function in aperture
(real symmetrical).In such cases, transmission function is absorbedOrderFormula (5) can be reduced to
Following expression, the i.e. Fourier spectrum of differential contrast image and quantitative phase system relation：
Therefore, it is rightCarry out inverse Fourier transform, you can obtain the quantitative phase information of the micro- test of sample, i.e., by micro-
The frequency-domain function of point phase contrast image can draw the quantitative phase information of the micro- test of sample.
Such as Fig. 5, divided with the single symmetry axis of any angle and obtain differential contrast imaging results as on+90 ° of directions of sample α
Phase contrast imaging result.
Because above-mentioned differential contrast imaging results are, with the differential contrast image of single symmetrical direction of principal axis acquisition, not wrap completely
Containing the phase information on symmetry axis；It is phase reconstruction ability on enhancing different directions, can be along any direction using many
Axle divides to form asymmetric lighting pattern, is imaged picture so as to obtain the differential contrast comprising different directions phase information, then enter
One step completes the differential contrast imaging containing out of phase information and is reduced to quantitative phase picture.Further, the present invention is used
Multi-direction symmetry axis divides to form asymmetric lighting pattern, and then using processing equipment to the target figure containing out of phase information
As being processed, specific processing method is as follows：
Step A：Obtain the differential contrast imaging picture that sample any two or multiple symmetrical direction of principal axis are divided
Definition is divided under illumination pattern with first symmetry axis, and the first object image of light collection is illuminated using the first difference
It is I11, the second target image for using the second difference to illuminate light collection is I12；Definition further divides photograph with j-th strip symmetry axis
Under bright pattern, the first object image for using the first difference to illuminate light collection is Ij1, using the of the second difference illumination light collection
Two target images are Ij2；Then differential contrast image is：
Step B：Differential contrast is set up based on weak signal target transmission function and is imaged two-dimension optical phase transfer function H (u), its table
It is as follows up to formula：
H (u)=i [∫ ∫ S (u ') P*(u′)p(u′+u)d2u′-∫∫S(u′)P*(u′)p(u′-u)d2u′]
Wherein, S (u ') is light source intensity distribution function, and P (u ') is pupil transmission function, and u ' is coordinate in pupil plane；
Step C：Set up differential contrast image in Fourier as follows with phase information relational model：
Aforesaid equation is combined and is solved, then calculated by equation below based on the quantitative phase information that multiaxis is divided：
Due to H (u) and the Frequency point axially along intersection be zero and participate in computing frequency it is wider, therefore directly deconvolution
The noise in these frequencies can be significantly superimposed, it is possible to which, by regularization least square method solve problem, final abbreviation is quantified
Phase information expression formula is as follows：Wherein, α is regularization parameter
Embodiments of the invention have been described in detail above, but the content is only presently preferred embodiments of the present invention,
And without of the invention with limitation.All any modifications made in application range of the invention, equivalent and improvement etc. all should
It is included within protection scope of the present invention.
Claims (10)
1. a kind of method based on differential contrast image restoring quantitative phase images, it is characterised in that comprise the following steps：
Step A：Using the LED array of programming Control as light source, based on time division multiplexing control LED array light source illumination figure
The switching of sample gathers the Frequency Synchronization of image with camera installation, and wherein target image is adopted under asymmetric lighting pattern
Collection, the asymmetric lighting pattern is to divide to form with single symmetry axis；Define the first mesh gathered under the first difference illumination light
Logo image is I1, the second target image under the second difference illumination light is I2, then differential contrast image is as follows：
Step B：Differential contrast is set up based on weak signal target transmission function and is imaged two-dimension optical phase transfer function H (u), its expression formula
It is as follows：
H (u)=i [∫ ∫ S (u ') P*(u′)p(u′+u)d2u′-∫∫S(u′)P*(u′)p(u′-u)d2u′]
Wherein, S (u ') is light source intensity distribution function, and P (u ') is pupil transmission function, and u ' is coordinate in pupil plane；
Step C：Set up differential contrast image in Fourier as follows with phase information relational model：
Then the quantitative phase information of reduction is on FourierIt is final rightInverse Fourier transform is carried out, is just obtained
Obtained quantitative phase information.
2. a kind of method based on differential contrast image restoring quantitative phase images, it is characterised in that comprise the following steps：
Step A：Using the LED array of programming Control as light source, based on time division multiplexing control LED array light source illumination figure
The switching of sample gathers the Frequency Synchronization of image with camera installation, and wherein target image is adopted under asymmetric lighting pattern
Collection, the asymmetric lighting pattern is to divide to form with the symmetry axis of two or more any directions；Collection target image is specific
Step operation in the following order：
A1：Illumination pattern is divided using first symmetry axis, defining the first object image gathered under the first difference illumination light is
I11, the second target image of gained is I under the second difference illumination light of collection12；
A2：Illumination pattern is further divided using j-th strip symmetry axis, the first object figure gathered under the first difference illumination light is defined
As being Ij1, the second target image gathered under the second difference illumination light is Ij2；
A3：The operation of repeat step A2 until reach the target number of axle, then the differential contrast of two width target images of each step gained
Image is as follows：
Step B：Differential contrast is set up based on weak signal target transmission function and is imaged two-dimension optical phase transfer function H (u)：
H (u)=i [∫ ∫ S (u ') P*(u′)p(u′+u)d2u′-∫∫s(u′)P*(u′)p(u′-u)d2u′]
Wherein, S (u ') is light source intensity distribution function, and P (u ') is pupil transmission function, and u ' is coordinate in pupil plane；
Step C：Set up differential contrast image in Fourier as follows with phase information relational model：
Aforesaid equation is combined and is solved, then calculated by equation below based on the quantitative phase information that multiaxis is divided：
3. a kind of method based on differential contrast image restoring quantitative phase images according to claim 1 and 2, its feature
It is that the LED array is the rectangular array being equidistantly arranged to make up by m × n light emitting diode, m, n is positive integer；
Lighting for LED array illumination pattern uses static screen pattern.
4. a kind of method based on differential contrast image restoring quantitative phase images according to claim 1 and 2, its feature
It is that the asymmetric lighting pattern is semicircle or semi-circular with LED array center as the center of circle or with LED array center
Symmetrical rectangle, illumination pattern is the first illumination pattern and the second illumination pattern, and first illumination pattern and described second are shone
Central Symmetry of the bright pattern on LED array, and the first illumination pattern and the second illumination pattern are while light to be formed with LED array
Center is circular, the annular or rectangle at midpoint.
5. a kind of method based on differential contrast image restoring quantitative phase images according to claim 1 and 2, its feature
It is, the also position correction including LED array light source before building LED array light source micro imaging system and carrying out IMAQ,
Specifically include LED array center be aligned with micro objective center, LED array horizontality calibration, LED array azimuth calibration.
6. a kind of quantitative phase reduces micro imaging system, it is characterised in that including transmission electron microscope, it is characterised in that institute
Transmission electron microscope light source is stated for LED array, the illumination pattern of the LED array passes through programming Control, also including for correcting
The multi-dimensional displacement platform of LED array light source position, gather microscopic result camera installation and by collection image restoring be
The processing equipment of quantitative phase images.
7. a kind of quantitative phase according to claim 6 reduces micro imaging system, it is characterised in that the LED array
It is the rectangular array being equidistantly arranged to make up by m × n light emitting diode, m, n is positive integer；LED array illumination pattern
Light using static screen pattern.
8. a kind of quantitative phase reduction micro imaging system according to claim 6, it is characterised in that processing equipment according to
The target image of collection obtains the quantitative phase imaging picture of sample, and its specific method is：
Step A：Obtain the differential contrast imaging picture I that sample is divided with any one symmetrical direction of principal axisDPC；Define the first difference
The first object image gathered under illumination light is I1, the second target image under the second difference illumination light is I2, then IDPCAcquisition
Method is：
Step B：Differential contrast is set up based on weak signal target transmission function and is imaged two-dimension optical phase transfer function H (u), its expression formula
It is as follows：
H (u)=i [∫ ∫ S (u ') P*(u′)p(u′+u)d2u′-∫∫S(u′)P*(u′)p(u′-u)d2u′]
Wherein, S (u ') is light source intensity distribution function, and P (u ') is pupil transmission function, and u ' is coordinate in pupil plane；
Step C：Set up differential contrast image in Fourier as follows with phase information relational model：
Then the quantitative phase information of reduction is on FourierIt is rightInverse Fourier transform is carried out, it is final to obtain
Quantitative phase information.
9. a kind of quantitative phase reduction micro imaging system according to claim 6, it is characterised in that processing equipment according to
The target image of collection obtains quantitative phase imaging picture, and its specific method is：
Step A：Obtain the differential contrast imaging picture that sample any two or multiple symmetrical direction of principal axis are divided
Definition is divided under illumination pattern with first symmetry axis, use the first difference illuminate the first object image of light collection for
I11, the second target image for using the second difference to illuminate light collection is I12；Definition further divides illumination with j-th strip symmetry axis
Under pattern, the first object image for using the first difference to illuminate light collection is Ij1, using the second of the second difference illumination light collection
Target image is Ij2；Then differential contrast image is：
Step B：Differential contrast is set up based on weak signal target transmission function and is imaged two-dimension optical phase transfer function H (u), its expression formula
It is as follows：
H (u)=i [∫ ∫ s (u ') P*(u′)p(u′+u)d2u′-∫∫s(u′)P*(u′)p(u′-u)d2u′]
Wherein, S (u ') is light source intensity distribution function, and P (u ') is pupil transmission function, and u ' is coordinate in pupil plane；
Step C：Set up differential contrast image in Fourier as follows with phase information relational model：
Aforesaid equation is combined and is solved, then calculated by equation below based on the quantitative phase information that multiaxis is divided：
10. a kind of quantitative phase according to claim 6~9 any one reduces micro imaging system, it is characterised in that
Also include controller, the controller is connected to control the multi-mode of LED array to shine respectively with LED array light source and camera installation
Bright pattern switching and camera installation gather the Frequency Synchronization of image.
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Cited By (14)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107272178A (en) * | 2017-08-04 | 2017-10-20 | 南京理工大学 | Differential phase contrast micro imaging method based on semicircular LED illumination |
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| CN108537842A (en) * | 2017-12-29 | 2018-09-14 | 南京理工大学 | The heteropical correction of background and compensation method in differential phase contrast micro-imaging |
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| CN111123497A (en) * | 2020-01-21 | 2020-05-08 | 电子科技大学 | Microscope optical device based on LED array illumination and calibration method thereof |
| WO2020103466A1 (en) * | 2018-11-22 | 2020-05-28 | 南京理工大学 | Phase contrast and differential interference contrast micro-imaging method based on light intensity transmission equation |
| CN112634800A (en) * | 2020-12-22 | 2021-04-09 | 北方液晶工程研究开发中心 | Method and system for rapidly and automatically testing refresh frequency of light-emitting diode display screen |
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| WO2022062773A1 (en) * | 2020-09-24 | 2022-03-31 | 南京理工大学 | Miniaturized, low-cost, and multi-contrast unmarked microscopic imaging system |
| CN114509869A (en) * | 2021-12-31 | 2022-05-17 | 南京理工大学智能计算成像研究院有限公司 | Multi-radius annular illumination-based microscopic imaging device and deconvolution method |
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104765138A (en) * | 2015-04-17 | 2015-07-08 | 南京理工大学 | Multi-mode micro-imaging system and method based on LED array |
| CN105158887A (en) * | 2015-09-29 | 2015-12-16 | 南京理工大学 | Multi-mode microimaging method based on programmable LED array illumination |
| CN105403988A (en) * | 2015-09-29 | 2016-03-16 | 南京理工大学 | Programmable aperture microscope system based on LCD liquid crystal panel and multi-mode imaging method thereof |
| CN105425377A (en) * | 2015-12-29 | 2016-03-23 | 电子科技大学 | Imaging method and system for microscope |
-
2016
- 2016-12-23 CN CN201611213588.4A patent/CN106768396A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104765138A (en) * | 2015-04-17 | 2015-07-08 | 南京理工大学 | Multi-mode micro-imaging system and method based on LED array |
| CN105158887A (en) * | 2015-09-29 | 2015-12-16 | 南京理工大学 | Multi-mode microimaging method based on programmable LED array illumination |
| CN105403988A (en) * | 2015-09-29 | 2016-03-16 | 南京理工大学 | Programmable aperture microscope system based on LCD liquid crystal panel and multi-mode imaging method thereof |
| CN105425377A (en) * | 2015-12-29 | 2016-03-23 | 电子科技大学 | Imaging method and system for microscope |
Non-Patent Citations (1)
| Title |
|---|
| LEI TIAN: "Quantitative differential phase contrast imaging in an LED array microscope", 《OPTICS EXPRESS》 * |
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