CN111260747B - High-flux optical tomography method and system based on virtual digital modulation - Google Patents

High-flux optical tomography method and system based on virtual digital modulation Download PDF

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CN111260747B
CN111260747B CN202010059012.7A CN202010059012A CN111260747B CN 111260747 B CN111260747 B CN 111260747B CN 202010059012 A CN202010059012 A CN 202010059012A CN 111260747 B CN111260747 B CN 111260747B
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袁菁
金锐
姜辰宇
龚辉
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Huazhong University of Science and Technology
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Abstract

The invention provides a high-flux optical tomography method based on virtual digital modulation, which comprises the steps of imaging a certain sampling area by N arrays to obtain N sampling images, virtually modulating the N sampling images, and demodulating to obtain a focal plane image of the sampling area. The sampling area is only required to be sampled once, and modulation is not carried out by means of a modulation device, so that the imaging speed is greatly increased; meanwhile, the virtual digital modulation can reduce noise introduced by the defocused signal by utilizing truncation integration, so that the imaging quality is better.

Description

High-flux optical tomography method and system based on virtual digital modulation
Technical Field
The invention relates to an optical imaging technology, in particular to a high-flux optical tomography method, an imaging system and a three-dimensional imaging system based on virtual digital modulation.
Background
The high-flux optical tomography method has great significance for biomedical research, and strong background fluorescence exists in thick tissue imaging of the traditional wide-field microscope. To solve this problem, several technical methods have been proposed, including confocal microscopy, two-photon microscopy, light-sheet microscopy, and structured light microscopy, among others. In the confocal microscope imaging technology, a small hole is arranged in front of a camera to block out-of-focus background signals, and only focal plane signals pass through, so that the optical tomography effect is realized. The two-photon microscopic imaging technology utilizes the nonlinear effect, and only has enough energy at the focal position to excite fluorescence, thereby having natural chromatographic effect. Both imaging methods belong to a point scanning imaging technology, and are low in imaging flux and difficult to be suitable for large sample imaging. The light sheet microscopic imaging adopts a mode of orthogonal illumination and detection, and only signals of an illuminated area of a light sheet can be received by a detection light path, so that the optical tomography effect is realized. The imaging flux of the light sheet imaging technology is high, but the light transparent technology is usually required to be combined, the processing flow is complex, and the problem of inconsistent spatial resolution exists due to the effect of factors such as scattering and the like.
The structured light microscopic imaging technology superposes a high-frequency periodic pattern on wide-field illumination to realize the modulation of a focal plane signal, and a defocusing signal is inhibited due to the rapid attenuation of the high-frequency modulation, so that the optical tomography is realized. The traditional structured light microscopic imaging is realized by a structured illumination and wide field imaging method, and compared with a point scanning mode of confocal microscopic imaging and other methods, the imaging speed is greatly improved. The chinese patent with application number 201210402820.4 discloses a method for rapid three-dimensional microscopic imaging of a large sample, which adopts a digital micromirror array to realize rapid structural light modulation, and realizes three-dimensional continuous imaging of the large sample through mosaic scanning and splicing. The mosaic scanning mode adopted by the method consumes a large amount of time on the movement of the translation stage, and meanwhile, the structured light modulation method based on the digital micromirror array can be used for carrying out structured light analytic reconstruction only by shooting at least 3 images in one field of view. In addition, the imaging quality of the imaging method has large dependence on modulation depth, and particularly when a thick tissue sample is imaged, strong background fluorescence causes the light modulation depth of the structure to be reduced, so that the signal-to-noise ratio of a reconstructed image is reduced. Therefore, there is a need for a simple and efficient high throughput optical tomography method and system.
Disclosure of Invention
The invention aims to overcome the technical defects, provides a high-flux optical tomography method and an imaging system based on virtual digital modulation, and solves the technical problems that the imaging speed of a structured light illumination microscopic imaging technology on a large-size sample is low, an additional modulation device is required to be used, and the dependence on the contrast of a modulation pattern is large in the prior art.
In order to achieve the technical purpose, the technical scheme of the invention provides a high-flux optical tomography method based on virtual digital modulation, which comprises the following steps:
s1, imaging a sample sampling area under a line illumination light spot by using a camera with N linear arrays to form original N sampling images p (i), wherein i represents the ith linear array of the camera, p (i) represents an original image of the sampling area acquired by the ith linear array of the camera, i and N are positive integers, and i is less than or equal to N;
s2, digitally modulating and integrating and summing all the acquired original images p (i) of the sampling area to generate a digital structured light image p of the sampling area at j phase j Comprises the following steps:
Figure BDA0002373795650000021
wherein m is j (i) The digital modulation function m (i) under the j phase is shown, the digital modulation function m (i) is a periodic function, the period is marked as T, and the m (i) meets the following conditions:
m(i)＝0 or 1,1≤i≤N
mod(N,T)＝0,T≥3
namely, the value of the periodic function m (i) is 0 or 1, the effective linear array number N of the linear array camera is integral multiple of the period T of m (i), and the period T of the periodic function is not less than 3;
s3, for at least three digital structured light images p with uniform phase distribution j Demodulating to obtain a focal plane image p in I.e. an optical tomographic image, a digital structured-light image p j Can be expressed in the following form:
p j ＝p in s j (x)+p out
Figure BDA0002373795650000022
wherein s is j (x) For digitally structured light images p j The intensity distribution of the corresponding modulation is,
Figure BDA0002373795650000023
is s is j (x) M is the modulation depth, T is the period of the digital modulation function M (i), x denotes the position of the sample, p in For sampling focal plane images of the image, p out An out-of-focus plane image which is a sampled image; focal plane image p in The demodulation algorithm of (a) is of the form:
Figure BDA0002373795650000024
Figure BDA0002373795650000031
wherein, | | denotes taking a modulus in the complex number domain, inum is a complex unit, C is a constant greater than 0, and J is the total number of modulation functions.
The invention provides a high-flux optical tomography method based on virtual digital modulation, which comprises the steps of imaging a certain sampling area by N arrays to obtain N sampling images, then virtually modulating the N sampling images, and demodulating to obtain a focal plane image of the sampling area. The sampling area is only required to be sampled once, and modulation is not carried out by means of a modulation device, so that the imaging speed is greatly increased; meanwhile, the virtual digital modulation can reduce the noise introduced by the defocused signal by utilizing 'truncation' integration, so that the imaging quality is better.
Compared with the prior art, the high-flux optical tomography method based on virtual digital modulation provided by the invention omits a physical modulation device, and meanwhile, has deeper imaging depth, better background suppression capability and higher image signal-to-noise ratio.
Meanwhile, the invention also provides a high-flux optical tomography system based on virtual digital modulation, which comprises:
the image acquisition unit is used for imaging a sample sampling area under a line illumination light spot by adopting a camera with N linear arrays to form original N sampling images p (i), wherein i represents the ith linear array of the camera, p (i) represents an original image of the sampling area acquired by the ith linear array of the camera, i and N are positive integers, and i is less than or equal to N;
an image modulation unit for performing digital modulation and integral summation on all the acquired original images p (i) of the sampling area to generate a digital structured light image p of the sampling area under j phase j Comprises the following steps:
Figure BDA0002373795650000032
wherein m is j (i) The digital modulation function m (i) under the j phase is shown, the digital modulation function m (i) is a periodic function, the period is marked as T, and the m (i) meets the following conditions:
m(i)＝0 or 1,1≤i≤N
mod(N,T)＝0,T≥3
namely, the value of the periodic function m (i) is 0 or 1, the effective linear array number of the linear array camera is integral multiple of the period of m (i), and the period of the periodic function is not less than 3;
an image demodulation unit for demodulating at least three digital structured light images p with uniform phase distribution j Demodulating to obtain a focal plane image p in I.e. an optical tomographic image, a digital structured-light image p j Can be expressed in the following form:
p j ＝p in s j (x)+p out
Figure BDA0002373795650000041
wherein s is j (x) For digitally structured light images p j The intensity distribution of the corresponding modulation is,
Figure BDA0002373795650000042
is s is j (x) M is the modulation depth, T is the period of the digital modulation function M (i), x denotes the position of the sample, p in For sampling focal plane images of the image, p out For separating sampled imagesA focal plane image; focal plane image p in The demodulation algorithm of (a) is of the form:
Figure BDA0002373795650000043
Figure BDA0002373795650000044
wherein, | | denotes taking a modulus in the complex number domain, inum is a complex unit, C is a constant greater than 0, and J is the total number of modulation functions.
The high-flux optical tomography system based on the virtual digital modulation is used for realizing a high-energy optical tomography method based on the virtual digital modulation.
The invention further provides a high-throughput optical tomography three-dimensional imaging device based on virtual digital modulation, which comprises the high-throughput optical tomography system based on virtual digital modulation, and further comprises: the object stage is used for placing an object to be imaged; and the cutting module is used for cutting off the object shallow layer after imaging is finished, and the sample is part of the object shallow layer.
The three-dimensional imaging device can perform tomography on the three-dimensional object, cut the shallow layer of the obtained focal plane image, and simultaneously acquire the corresponding focal plane image of the newly exposed shallow layer of the object by adopting the high-energy optical tomography method based on virtual digital modulation, and cut the focal plane image, thereby reducing the imaging time of the three-dimensional object.
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FIG. 1 is a reconstruction diagram of the high throughput optical tomography method of the present invention.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is described in further detail below with reference to the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.
As shown in FIG. 1, the present invention provides a high throughput optical tomography method based on virtual digital modulation, comprising the following steps:
s1, as shown in fig. 1 (a), imaging a sample sampling region under a line illumination spot by using a camera having N linear arrays, where i is a positive integer and is not less than 1 and not more than N, and p (i) represents an original image of the sampling region obtained by the ith linear array of the camera, that is, obtaining sampling images p (1) -p (N); when in specific imaging, the method comprises the following steps:
s101, as shown in (b) of FIG. 1, defining a region, where a sample is located at the maximum light intensity of a line illumination light spot, as a sample sampling region, and imaging the sample sampling region;
s102, moving a camera and imaging a sample, wherein the imaging single-frame exposure time is the same as the time for moving the sample by a line array;
and S103, finishing imaging when the sample moves out of the position where the light intensity of the line illumination light spot is strongest.
In this embodiment, the light beams are shaped into linear illumination light spots, and then the linear illumination light spots are focused on the focal plane position of the objective lens, the thickness of the light at the focal plane position of the objective lens is limited by the optical diffraction law, and theoretically, the thinner the linear light beam is, the larger the modulation depth is, and the better the signal-to-noise ratio of the structured light reconstructed image is. The diffraction-limited line illumination light spot can be perpendicular to the moving direction of the sample, the embodiment can drive the sample to move continuously and uniformly along the direction perpendicular to the diffraction-limited line beam, and can also drive the diffraction-limited line beam to move continuously and uniformly along the direction parallel to the sample, as long as the diffraction-limited line beam and the sample can generate relative continuous and uniform motion.
As shown in fig. 1 (a), the camera of the present embodiment is an N-line area array detector, and thus includes N lines of linear arrays; perpendicular X and Y directions are formed on a plane parallel to the imaging plane of the sample. And the distribution direction and the width of the N-line array are respectively the same as those of the diffraction-limited line illumination light spots and are in conjugate relation with each other, so that the imaging area is convenient to correspond to the line illumination modulation light beam. The moving direction of the sample relative to the diffraction-limited line illuminating light spot is also along the X direction, so that the moving direction of the sample relative to the diffraction-limited line light beam is ensured to be the same as the arrangement direction of the N rows of linear arrays.
S2, as shown in (c) of fig. 1, digitally modulating and integrating the original images obtained from the N linear arrays of the camera, and generating the j-th phase digital structured light image expression as follows:
Figure BDA0002373795650000051
wherein m is j (i) The digital modulation function m (i) under the j phase is shown, the digital modulation function m (i) is a periodic function, the period is marked as T, and the m (i) meets the following conditions:
m(i)＝0 or 1,1≤i≤N
mod(N,T)＝0,T≥3
namely, the value of the periodic function m (x) is 0 or 1, and the effective line number N of the line camera is an integral multiple of the period T of m (x). Theoretically, infinite integration is required for the detection surface, and the number of linear arrays of an actual camera is limited, so that truncation integration is required. Furthermore, truncation of the integration removes part of the out-of-focus signal, which helps to improve the signal-to-noise ratio of the reconstructed image, for reasons that will be briefly explained below.
The structured light microscopic imaging technology superposes high-frequency periodic patterns on wide-field illumination to realize the modulation of focal plane signals, and defocusing signals are inhibited due to the rapid attenuation of high-frequency modulation, so that optical tomography is realized. However, noise is inevitably present in the actual imaging, and the noise of the defocus signal also affects the signal-to-noise ratio of the reconstructed image, and the signal-to-noise ratio of the structured light tomography image can be expressed as follows:
Figure BDA0002373795650000061
wherein the content of the first and second substances,<>representing the mathematical expectation, var () represents the variance. As can be seen from this equation, the defocus signal p out The variance of (i.e., noise) can affect the signal-to-noise ratio of the reconstructed image. The truncation integration of the scheme can be understood as the action of a 'small hole' in confocal imaging, and the 'small hole' removes part of the defocused signal, so that the defocused signal p is reduced out The variance of (i.e., noise) and thus the signal-to-noise ratio of the reconstructed image. It should be noted that the integration interval for truncation of the integration must be an integer number of modulation periods, otherwise the "energy leakage" problem may occur. In addition, classical structural optical tomography reconstruction requires at least 3 different phase images, the period is not less than 3 in order to generate at least 3 phases;
s3, continuing as shown in (c) of FIG. 1, for at least three digital structured light images p with uniform phase distribution j Demodulating to obtain a focal plane image p in I.e. an optical tomographic image, a digital structured-light image p j Expressed as:
p j ＝p in s j (x)+p out
Figure BDA0002373795650000062
wherein s is j (x) For digitally structured light images p j The intensity distribution of the corresponding modulation is,
Figure BDA0002373795650000063
is s is j (x) M is the modulation depth, T is the period of the digital modulation function M (i), x denotes the position of the sample, p in For sampling focal plane images of the image, p out An out-of-focus plane image, focal plane image p, of the sample image in The demodulation algorithm of (a) is of the form:
Figure BDA0002373795650000064
Figure BDA0002373795650000071
wherein, | | denotes taking a modulus in the complex number domain, inum is a complex unit, C is a constant greater than 0, and J is the total number of modulation functions.
Aiming at the imaging method, the invention also provides a high-flux optical tomography system based on virtual digital modulation, which comprises the following steps:
the image acquisition unit is used for imaging a sample sampling area under a line illumination light spot by adopting a camera with N linear arrays to form original N sampling images p (i), wherein i represents the ith linear array of the camera, p (i) represents an original image of the sampling area acquired by the ith linear array of the camera, i and N are positive integers, and i is less than or equal to N;
an image modulation unit for performing digital modulation and integral summation on all the acquired original images p (i) of the sampling area to generate a digital structured light image p of the sampling area under j phase j Comprises the following steps:
Figure BDA0002373795650000072
wherein m is j (i) The digital modulation function m (i) under the j phase is shown, the digital modulation function m (i) is a periodic function, the period is marked as T, and the m (i) meets the following conditions:
m(i)＝0 or 1,1≤i≤N
mod(N,T)＝0,T≥3
the value of a periodic function m (i) is 0 or 1, the effective linear array number N of the linear array camera is an integral multiple of the period T of m (i), and the period T of the periodic function is not less than 3;
an image demodulation unit for demodulating at least three digital structured light images p with uniform phase distribution j Demodulating to obtain a focal plane image p in I.e. an optical tomographic image, a digital structured-light image p j Can be expressed in the following form:
p j ＝p in s j (x)+p out
Figure BDA0002373795650000073
wherein s is j (x) For digitally structured light images p j The intensity distribution of the corresponding modulation is,
Figure BDA0002373795650000074
is s is j (x) M is the modulation depth, T is the period of the digital modulation function M (i), x denotes the position of the sample, p in For sampling focal plane images of the image, p out An out-of-focus plane image which is a sampled image; focal plane image p in The demodulation algorithm of (a) is of the form:
Figure BDA0002373795650000075
Figure BDA0002373795650000081
wherein, | | denotes taking a modulus in the complex number domain, inum is a complex unit, C is a constant greater than 0, and J is the total number of modulation functions.
The invention provides a high-flux optical tomography method and system based on virtual digital modulation, which are characterized in that N arrays are imaged on a certain sampling area to obtain N sampling images, and then the N sampling images are subjected to virtual modulation and demodulation to obtain a focal plane image of the sampling area. The sampling area is only required to be sampled once, and modulation is not carried out by means of a modulation device, so that the imaging speed is greatly increased; meanwhile, when virtual digital modulation is carried out, the noise introduced by the defocused signal can be reduced by utilizing truncation integration, so that the imaging quality is better.
The imaging method is suitable for imaging a single sheet sample and also suitable for imaging a three-dimensional object, and the imaging of the single sheet sample is taken as an example for explanation.
Example 1
The method comprises the steps of photographing a sample by using a camera with a line number N being 4, namely, a single-frame image comprises 4 rows of pixels p (1) -p (4), and according to the condition that the effective line number N of the line camera is an integral multiple of a period T of a digital modulation function m (i), at least three different phase images are required for the reconstruction of the structured light image, so that the period T of the digital modulation function m (i) is 4.
In order to obtain a structured light modulation image with uniform phase distribution, 4 different phase images need to be generated through virtual digital modulation, so the values of the digital modulation functions m (i) of 4 different phases in the period T ═ 4 can be: 1100, 0110, 0011 and 1001. According to the digital modulation method, 4 different phase images can be calculated as follows:
p 1 ＝p(1)+p(2)
p 2 ＝p(2)+p(3)
p 3 ＝p(3)+p(4)
p 4 ＝p(4)+p(1)
wherein p is 1 ～p 4 Is recorded as
Figure BDA0002373795650000086
The phase interval is pi/2. Here, it is to be noted that p 1 ～p 4 Phase of
Figure BDA0002373795650000082
The specific value of (a) theoretically depends on the digital modulation function m (i). However, since
Figure BDA0002373795650000083
Is 4 phases which are uniformly distributed, so that the actual calculation can be carried out
Figure BDA0002373795650000084
Simply take the values 0, pi/2, pi, 3 pi/2. According to the structured light tomography principle, 4 structured light images p with uniform phase distribution 1 ～p 4 Expressed as:
p 1 ＝p in s 1 (x)+p out ，
Figure BDA0002373795650000085
p 2 ＝p in s 2 (x)+p out ，
Figure BDA0002373795650000091
p 3 ＝p in s 3 (x)+p out ，
Figure BDA0002373795650000092
p 4 ＝p in s 4 (x)+p out ，
Figure BDA0002373795650000093
it should be noted that, in practice, the value of x may be different, but in the above formula, the value does not play a role, and therefore, can be disregarded;
focusing a planar image p in combination with the above formula in And (3) demodulation:
Figure BDA0002373795650000094
in the formula (I), originally
Figure BDA0002373795650000095
M is the modulation depth and J is the total number of modulation functions, but the constant C is generally negligible in practical calculations. And p (1) -p (4) in the finally obtained formula come from a single-frame image shot by a camera, so that a focal plane image can be demodulated. In actual imaging, if a translation stage is adopted to drive a sample to move, a sample signal sequentially enters the central position of a diffraction limited line light spot, and each sampling area corresponds to one frame of image shot by a camera and can be demodulated according to the formula.
Example 2
The method comprises the steps of photographing a sample by using a camera with the line number N being 6, namely, a single-frame image comprises 6 lines of pixels p (1) -p (6), and according to the condition that the effective line number N of the line camera is an integral multiple of a period T of a digital modulation function m (i), at least three different phase images are required for the reconstruction of the structured light image, wherein the period T of the digital modulation function m (i) is 6.
In order to obtain a structured light modulation image with uniform phase distribution, 3 different phase images need to be generated through virtual digital modulation, so the values of the digital modulation functions m (i) of 3 different phases in the period T ═ 6 can be: 111000, 001110 and 100011. According to the digital modulation method, 3 different phase images can be calculated as follows:
p 1 ＝p(1)+p(2)+p(3)
p 2 ＝p(3)+p(4)+p(5)
p 3 ＝p(5)+p(6)+p(1)
wherein p is 1 ～p 3 Is recorded as
Figure BDA0002373795650000097
The phase interval is 2 pi/3. Here, it is to be noted that p 1 ～p 3 Phase of
Figure BDA0002373795650000096
The specific value of (a) theoretically depends on the digital modulation function m (i). However, since
Figure BDA0002373795650000101
Is 3 phases which are uniformly distributed, so that the actual calculation can be carried out
Figure BDA0002373795650000102
Simply take the values 0, 2 pi/3, 4 pi/3. According to the structured light tomography principle, 3 structured light images p with uniform phase distribution 1 ～p 3 Expressed as:
p 1 ＝p in s 1 (x)+p out ，
Figure BDA0002373795650000103
p 2 ＝p in s 2 (x)+p out ，
Figure BDA0002373795650000104
p 3 ＝p in s 3 (x)+p out ，
Figure BDA0002373795650000105
it should be noted that, in practice, the value of x may be different, but in the above formula, the value does not play a role, and therefore, can be disregarded;
focusing a planar image p in combination with the above formula in And (3) demodulation:
Figure BDA0002373795650000106
in the formula (I), originally
Figure BDA0002373795650000107
M is the modulation depth and J is the total number of modulation functions, but the constant C is generally negligible in practical calculations. And p (1) -p (6) in the finally obtained formula come from a single-frame image shot by a camera, so that a focal plane image can be demodulated. In actual imaging, if a translation stage is adopted to drive a sample to move, a sample signal sequentially enters the central position of a diffraction limited line light spot, and each sampling area corresponds to one frame of image shot by a camera and can be demodulated according to the formula.
The above 2 embodiments are described by taking a thin slice sample as an example, and actually, the imaging method and system are also applicable to imaging of a three-dimensional object. The following is a detailed description of imaging of three-dimensional objects.
Example 3
The invention also provides a virtual digital modulation-based high-throughput optical tomography three-dimensional imaging device, which comprises the virtual digital modulation-based high-throughput optical tomography system and further comprises:
the object stage is used for placing an object to be imaged;
and the cutting module is used for cutting off the object shallow layer after imaging is finished, and the sample is part of the object shallow layer.
The three-dimensional object and the thin slice imaging have a great difference that the imaging of the three-dimensional object needs to demodulate the sample by adopting the high-flux optical tomography system based on the virtual digital modulation on the shallow layer, then the imaged shallow layer needs to be cut off to expose the new shallow layer, the sample is continuously demodulated by adopting the high-flux optical tomography system based on the virtual digital modulation, and the imaging-cutting-imaging mode is circularly adopted until all imaging of the whole three-dimensional object is completed.
In the imaging process, in order to save imaging time, except for the first cutting and the last imaging, the imaging system completes demodulation of a focal plane image of a shallow layer of an object cut last time while the cutting is performed. Therefore, the imaging can be carried out while cutting, and the imaging time is saved.
Further, the cutting module and the imaging object move relatively to realize that the cutting module cuts off the shallow layer of the object after imaging is finished. Whether the cutting module moves or the imaging object moves, the cutting can be realized as long as the cutting module and the imaging object move relatively.
Further, the three-dimensional moving platform is connected with the object stage or the cutting module. By adopting the three-dimensional moving platform, on one hand, the moving automation can be realized, and on the other hand, the position can be controlled more accurately.
The above-described embodiments of the present invention should not be construed as limiting the scope of the present invention. Any other corresponding changes and modifications made according to the technical idea of the present invention should be included in the protection scope of the claims of the present invention.

Claims (9)

1. A high-flux optical tomography method based on virtual digital modulation is characterized by comprising the following steps:
s1, imaging the sample sampling area under the line illumination spot by using a camera with N linear arrays to form original N sampling images p (i), wherein i represents the cameraThe i-th linear array of the camera represents an original image of a sampling area obtained by the i-th linear array of the camera, i and N are positive integers, i is less than or equal to N, the cameras of the N linear arrays respectively image the same sample sampling area at the same time, namely, each frame forms original N sampling images p (i) for the same sample sampling area; s2, digitally modulating and integrating and summing all the acquired original images p (i) of the sampling area to generate a digital structured light image p of the sampling area at j phase j Comprises the following steps:
Figure FDA0003654399150000011
the digital structured light image p j Representing images of different phases, in which m j (i) The digital modulation function m (i) under the j phase is shown, the digital modulation function m (i) is a periodic function, the period is marked as T, and the m (i) meets the following conditions:
m(i)＝0 or 1,1≤i≤N
mod(N,T)＝0,T≥3
namely, the value of the periodic function m (i) is 0 or 1, the effective linear array number N of the linear array camera is integral multiple of the period T of m (i), and the period T of the periodic function is not less than 3;
s3, for at least three digital structured light images p with uniform phase distribution j Demodulating to obtain focal plane image p in I.e. an optical tomographic image, a digital structured-light image p j Expressed in the following form:
p j ＝p in s j (x)+p out
Figure FDA0003654399150000012
wherein s is j (x) For digitally structured light images p j The intensity distribution of the corresponding modulation is,
Figure FDA0003654399150000013
is s is j (x) M is the modulation depth,t is the period of the digital modulation function m (i), x denotes the position of the sample, p in For sampling focal plane images of the image, p out For out-of-focus plane images of the sampled image, focal plane image p in The demodulation algorithm of (a) is of the form:
Figure FDA0003654399150000014
Figure FDA0003654399150000021
wherein, | | denotes taking a modulus in the complex number domain, inum is a complex unit, C is a constant greater than 0, and J is the total number of modulation functions.
2. The virtual digital modulation-based high-throughput optical tomography method of claim 1, wherein the specific method for imaging the sample sampling region under the line illumination light spot by using the camera with N linear arrays in step S1 is as follows:
s101, defining a region, where a sample is located at the line illumination light spot and has the strongest light intensity, as a sample sampling region, and imaging the sample sampling region;
s102, moving a camera and imaging a sample, wherein the exposure time of a single imaging frame is the same as the time of moving the sample by a line array;
and S103, finishing imaging when the sample moves out of the position where the light intensity of the line illumination light spot is strongest.
3. The virtual digital modulation-based high throughput optical tomography method according to claim 2, wherein the at least three digital structured light images p j Successively corresponding modulated intensity distributions s j (x) Belonging to a uniform phase distribution.
4. The virtual digital modulation based high throughput optical tomography method of claim 1, wherein the digital modulation function m (i) is a square wave function.
5. A virtual digital modulation-based high throughput optical tomography system comprising:
the image acquisition unit is used for imaging a sample sampling area under a line illumination light spot by adopting a camera with N linear arrays to form original N sampling images p (i), wherein i represents the ith linear array of the camera, p (i) represents an original image of the sampling area obtained by the ith linear array of the camera, i and N are positive integers, i is less than or equal to N, the cameras with the N linear arrays respectively image the same sample sampling area at the same time, namely, each frame forms the original N sampling images p (i) to the same sample sampling area;
an image modulation unit for performing digital modulation and integral summation on all the acquired original images p (i) of the sampling area to generate a digital structured light image p of the sampling area under j phase j Comprises the following steps:
Figure FDA0003654399150000022
the digital structured light image p j Representing images of different phases, in which m j (i) The digital modulation function m (i) under the j phase is shown, the digital modulation function m (i) is a periodic function, the period is marked as T, and the m (i) meets the following conditions:
m(i)＝0 or 1,1≤i≤N
mod(N,T)＝0,T≥3
namely, the value of the periodic function m (i) is 0 or 1, the effective linear array number N of the linear array camera is integral multiple of the period T of m (i), and the period T of the periodic function is not less than 3;
an image demodulation unit for demodulating at least three digital structured light images p with uniform phase distribution j Demodulating to obtain focal plane image p in I.e. an optical tomographic image, a digital structured-light image p j Expressed in the following form:
p j ＝p in s j (x)+p out
Figure FDA0003654399150000031
wherein s is j (x) For digitally structured light images p j The intensity distribution of the corresponding modulation is,
Figure FDA0003654399150000032
is s is j (x) M is the modulation depth, T is the period of the digital modulation function M (i), x denotes the position of the sample, p in For sampling focal plane images of the image, p out An out-of-focus plane image which is a sampled image; focal plane image p in The demodulation algorithm of (a) is of the form:
Figure FDA0003654399150000033
Figure FDA0003654399150000034
wherein, | | denotes taking a modulus in the complex number domain, inum is a complex unit, C is a constant greater than 0, and J is the total number of modulation functions.
6. A virtual digital modulation-based high-throughput optical tomography three-dimensional imaging device, comprising the virtual digital modulation-based high-throughput optical tomography system of claim 5, further comprising:
the object stage is used for placing an object to be imaged;
and the cutting module is used for cutting off the object shallow layer after imaging is finished, and the sample is part of the object shallow layer.
7. The virtual digital modulation-based high-throughput optical tomography three-dimensional imaging device according to claim 6, wherein the imaging system completes demodulation of the focal plane image of the object shallow layer of the last cutting at the same time of the cutting except for the first cutting and the last imaging.
8. The virtual digital modulation-based high-throughput optical tomography three-dimensional imaging device according to claim 7, wherein the cutting module moves relative to the imaged object to realize the cutting module to cut off the shallow layer of the imaged object.
9. The virtual digital modulation-based high throughput optical tomography three-dimensional imaging apparatus according to claim 8, further comprising a three-dimensional moving platform connected to the stage or the cutting module.
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