Light field microscope based image acquisition method and apparatus

Abstract

The present invention is characterized by including a lenslet synthesis processing unit that acquires a virtual lenslet image using a group of multiple lenslet images, a subaperture image conversion unit that converts the virtual lenslet image generated by the lenslet synthesis processing unit into a subaperture image, and a 3D conversion unit that converts the subaperture image generated by the subaperture image conversion unit into a focal stack image.

Description

[Technical Field]

[0001] The present invention relates to a light field microscope-based image acquisition method and device that allows changes in a sample to be acquired as three-dimensional images with high spatial resolution. [Background technology]

[0002] Conventional fluorescence microscopes capture two-dimensional images of a sample, so to confirm the distribution of fluorescence in three-dimensional space, multiple slice images are captured in the z-axis direction and then combined to reconstruct the image in three dimensions.

[0003] That is, conventionally, a 3D image is obtained by precisely adjusting the focal length of the z-axis and combining images taken at each depth.

[0004] However, the above-mentioned method involves taking multiple images while changing the z-axis position, which can lead to distortion due to changes in the fluorescence of the sample during the imaging period. In addition, in the case of moving cell clusters (muscle cells), it is difficult to ascertain 3D information about the movement, making it extremely difficult to obtain an entire image at the same time.

[0005] A light field microscope was devised to solve this problem.

[0006] A light field microscope has an optical structure in which a lens array is added to the structure of a conventional fluorescence microscope, and images acquired from this optical structure can be converted into three-dimensional images using a restoration method.

[0007] However, while conventional light field microscopes have the advantage of being able to acquire 3D image information with a single shot, they have the disadvantage that the spatial resolution is lower than that of conventional microscopes because it depends on the number of lenses in the lens array.

[0008] That is, in the case of a light field microscope, there is a problem in that the resolution is significantly reduced due to the gaps between the lenses that make up the lens array.

[0009] Therefore, methods have been proposed to reduce the size of the lenses that make up the lens array, but this poses the problem of having to reduce the area of ​​the image sensor pixels.For reference, the area of ​​the image sensor pixels affects the sensitivity of the acquired image, so it is important to maintain a large pixel size for application to fluorescence imaging, which requires high sensitivity.

[0010] Therefore, the applicant has proposed the present invention in order to solve the above-mentioned problems, and a related prior art document is Korean Patent Registration No. 10-2253320 entitled "Method for displaying three-dimensional images in an integrated image microscope system and an integrated image microscope system realizing the same." Summary of the Invention [Problem to be solved by the invention]

[0011] The present invention has been made to solve these problems, and its purpose is to provide a light field microscope-based image acquisition method and apparatus that are configured to acquire images with high spatial resolution by reducing the size of the microlenses that make up the lens array, without increasing their number. [Means for solving the problem]

[0012] In order to achieve the above object, the present invention can include a lenslet synthesis processing unit that acquires a virtual lenslet image using a group of multiple lenslet images, a subaperture image conversion unit that converts the virtual lenslet image generated by the lenslet synthesis processing unit into a subaperture image, and a three-dimensional conversion unit that converts the subaperture image generated by the subaperture image conversion unit into a focal stack image.

[0013] The multiple lenslet images may be generated by acquiring a plurality of image frame information through microlenses whose arrangement positions are changed according to time periods within a set image sensor area.

[0014] The multiple lenslet images can be acquired by rotating a disk on which a plurality of microlenses are arranged for a set period of time.

[0015] The lenslet synthesis processor can also superimpose a plurality of image frames that constitute a group of multiple lenslet images to grasp the arrangement of microlenses for each image.

[0016] The sub-aperture image conversion unit may search for pixels corresponding to coordinates of the microlenses in the virtual lenslet image and map the pixels to coordinate positions of the sub-aperture image to be converted.

[0017] Further, the subaperture image conversion unit generates pixel values ​​of the virtual lenslet image using the following formula 1:

number

[0018] In addition, the 3D conversion unit converts the subaperture image into a focal stack image having 3D information on the x-axis, y-axis, and z-axis, and the x-axis and y-axis positions of the focal stack image can include 2D image information at the corresponding z-axis coordinate point.

[0019] Further, the three-dimensional conversion unit converts the sub-aperture image into a focal stack image using the following Equations 2 and 3:

number

number

[0020] The disk may also be coupled to an actuator or motor that provides rotational or linear power.

[0021] The linear movement range of the disk is 0 to 500 mm.

[0022] The method may also include a first step of acquiring a group of multiple lenslet images, a second step of acquiring a virtual lenslet image using the group of multiple lenslet images acquired in the first step, a third step of converting the virtual lenslet image acquired in the second step into a subaperture image, and a fourth step of converting the subaperture image converted in the third step into a focal stack image. [Effects of the Invention]

[0023] The light field microscope-based image acquisition method and apparatus of the present invention provides a configuration that can acquire a group of multiple lenslet images by rotating and linearly moving a disk on which microlenses are arranged, and generate a three-dimensional image using the group of multiple lenslet images, thereby making it possible to acquire three-dimensional images with higher spatial resolution than conventional light field microscopes.

[0024] Furthermore, the light field microscope-based image acquisition method and device according to the present invention enable three-dimensional image information of a moving measurement sample (such as a muscle cell) to be acquired more quickly and accurately than ever before.

[0025] In addition, the light field microscope-based image acquisition method and apparatus of the present invention can acquire images with high spatial resolution by preventing the phenomenon of reduced spatial resolution due to gaps between microlenses by changing the arrangement of the microlenses as the disk rotates.

[0026] Furthermore, the light field microscope-based image acquisition method and device of the present invention can acquire high spatial resolution images by reducing the size of the microlenses without increasing the number of microlenses arranged on the disk or reducing the size of the image sensor, thereby reducing the manufacturing costs of the device and significantly reducing the costs involved in acquiring images. [Brief explanation of the drawings]

[0027] [Figure 1] FIG. 1 is a block diagram illustrating the configuration of a light field microscope-based image acquisition device according to an embodiment of the present invention.

[0028] [Figure 2] 1 is a diagram illustrating a schematic configuration of a light field microscope to which an image acquisition device according to an embodiment of the present invention is applied.

[0029] [Figure 3] 3 is a diagram showing the path of light that passes through a disc according to an embodiment of the present invention and reaches an image sensor; FIG.

[0030] [Figure 4] 10A and 10B are diagrams illustrating a method for acquiring a plurality of images within a unit time by rotating a disk on which a plurality of microlenses are arranged for a unit time.

[0031] [Figure 5] FIG. 10 is a diagram showing how a group of multiple lenslet images is converted into a virtual lenslet image by a lenslet synthesis processor.

[0032] [Figure 6] FIG. 10 shows in more detail how multiple lenslet images are transformed into a virtual lenslet image.

[0033] [Figure 7] FIG. 1 illustrates a method for generating pixel values ​​in a virtual lenslet image.

[0034] [Figure 8] 10A and 10B are diagrams illustrating a processing procedure of a sub-aperture image conversion unit according to an embodiment of the present invention.

[0035] [Figure 9] FIG. 10 is a diagram showing the processing steps of a three-dimensional conversion unit that receives a sub-aperture image as an input.

[0036] [Figure 10] 1A to 1C are diagrams showing various arrangement patterns of microlenses provided on a disc.

[0037] [Figure 11] 1 is a flowchart of a light field microscope-based image acquisition method according to an embodiment of the present invention. DETAILED DESCRIPTION OF THE INVENTION

[0038] The advantages and features of the present invention, as well as the methods for achieving them, will become more apparent with reference to the following detailed embodiments in conjunction with the accompanying drawings.

[0039] However, the present invention is not limited to the embodiments disclosed below, but may be realized in various different forms, and the present embodiments are provided merely to complete the disclosure of the present invention and to fully convey the scope of the invention to those skilled in the art to which the present invention pertains. The present invention is defined only by the scope of the claims.

[0040] Hereinafter, a light field microscope-based image acquisition method and apparatus according to an embodiment of the present invention will be described in detail with reference to Figures 1 to 11. In describing the present invention, detailed descriptions of related well-known functions or configurations will be omitted to avoid obscuring the gist of the present invention.

[0041] FIG. 1 is a block diagram showing the configuration of a light field microscope-based image acquisition device according to one embodiment of the present invention. FIG. 2 is a diagram showing the configuration of a light field microscope to which an image acquisition device according to one embodiment of the present invention is applied. FIG. 3 is a diagram showing the path of light that passes through a disk according to one embodiment of the present invention until it reaches an image sensor. FIG. 4 is a diagram showing a method of acquiring multiple images within a unit time by rotating a disk on which multiple microlenses are arranged for a unit time. FIG. 5 is a diagram showing how a group of multiple lenslet images is converted into a virtual lenslet image by a lenslet synthesis processing unit. FIG. 6 is a diagram showing in detail the process of converting a group of multiple lenslet images into a virtual lenslet image. FIG. 7 is a diagram showing a method of generating pixel values ​​in a virtual lenslet image. FIG. 8 is a diagram showing the processing process of a subaperture image conversion unit according to one embodiment of the present invention. FIG. 9 is a diagram showing the processing process of a 3D conversion unit that inputs a subaperture image. FIG. 10 is a diagram showing various arrangement patterns of microlenses provided on a disk. FIG. 11 is a flowchart of a light field microscope-based image acquisition method according to one embodiment of the present invention.

[0042] As shown in Figures 1 and 2, a light field microscope-based image acquisition device 100 according to one embodiment of the present invention may include a lenslet synthesis processing unit 200 that acquires a virtual lenslet image using a multiple lenslet image group 10, a sub-aperture image conversion unit 300 that converts the virtual lenslet image generated by the lenslet synthesis processing unit 200 into a sub-aperture image, and a 3D conversion unit 400 that converts the sub-aperture image generated by the sub-aperture image conversion unit 300 into a focal-stack image.

[0043] The light field microscope-based image acquisition device 100 configured as described above can acquire a multi-lenslet image group 10 by rotating or linearly moving a disk D on which multiple microlenses ML are arranged, changing the arrangement position of the microlenses according to time periods, thereby acquiring multiple images.

[0044] For reference, FIG. 2 is a schematic illustration of a light field microscope to which a disk D according to an embodiment of the present invention is applied.

[0045] As shown in Figure 2, when a fluorescently stained measurement sample is irradiated with light of a specific wavelength that reacts with the fluorophore through a light source, fluorescent light of a specific wavelength is emitted from the measurement sample and can pass through the objective lens. The light that passes through the objective lens can then pass through a dichroic mirror. The wavelength of the light source is reflected by the dichroic mirror, and only the fluorescent wavelength passes through.

[0046] The fluorescent light that has passed through the dichroic mirror passes through a tube lens and an optical tube and reaches its focal plane, at which a disk D according to one embodiment of the present invention is placed.

[0047] The multiple microlenses ML installed on the disc D align the light that converges on the focal plane according to the position of each lens. The light that passes through the microlenses ML then passes through a relay lens. The relay lens adjusts the physical distance between the microlenses ML and the image sensor. The light that passes through the relay lens reaches the image sensor, which captures an image. The light path information captured through the image can be reconstructed into a 3D image using a software algorithm.

[0048] A part of the area of ​​the disk D may be disposed between the optical tube and the relay lens, and the center of the disk D may be connected to a drive unit M, which may be a known actuator or motor for transmitting rotational or linear power.

[0049] Therefore, the disk D is capable of linear reciprocating motion in the Z-axis direction shown in Fig. 2, and is also capable of rotational motion in the R direction shown in Fig. 2. For reference, the linear movement range of the disk D is preferably 0 to 500 mm. Of course, the rotational speed and linear movement speed of the disk D can be set to various values ​​depending on the measurement sample, measurement method, and image information to be obtained.

[0050] When the disk D rotates, the arrangement position of the microlenses ML is changed, so that different images can be acquired at different times depending on the changed arrangement position of the microlenses ML.

[0051] Therefore, as shown in FIGS. 4-6, the disk D can be rotated for a set time to acquire a set of multiple lenslet images 10.

[0052] That is, a plurality of image frame information is acquired by the microlenses ML, the arrangement position of which is changed according to time periods within a set image sensor area, and based on this information, a multi-lenslet image group 10 can be acquired. In this case, the method of changing the arrangement position of the microlenses ML can be realized by a structure in which a disk D on which a plurality of microlenses ML are arranged is rotated and linearly moved for a set time.

[0053] FIG. 4 shows a method for capturing a plurality of images within a unit time while rotating the disk D for that unit time.

[0054] As shown in FIG. 4, images are acquired from the image sensor while rotating a disk D on which microlenses ML are arranged according to time periods, and it can be seen that the arrangement positions of the corresponding microlenses ML are set differently for each image.

[0055] As described above, it is possible to obtain a multi-lenslet image group 10 by combining multiple image frame information acquired within a set time period through the array positions of the microlenses ML that are changed according to the time period.

[0056] For reference, Figure 3 shows the path of light that passes through the microlens ML before reaching the image sensor. The light that passes through the tube lens is focused at the position where the microlens ML is located, and the light that passes through the microlens ML spreads out again before passing through the relay lens. The light that passes through the relay lens is then focused again. The light information is collected and converted into a digital image signal by an image sensor at a position past the focus.

[0057] Although FIG. 4 shows that a plurality of microlenses ML are arranged on the disk D in a grid pattern, the present invention is not limited to this.

[0058] For example, as shown in FIG. 10, the microlenses ML can be arranged on the disk in a Nipkow pattern, a honeycomb pattern, or a random pattern.

[0059]

[0060] The lenslet synthesis processor 200 can grasp the arrangement of microlenses for each image by superimposing a plurality of image frames that constitute the multiple lenslet image group 10 .

[0061] As shown in Fig. 5, a virtual lenslet image can be acquired by inputting a multiple lenslet image group 10 consisting of multiple lenslet images acquired within a unit time into a lenslet synthesis processor 200. Fig. 5 shows a virtual lenslet image acquired using six lenslet images, but the number of lenslet images is not limited to six, and it is of course possible to adjust the number of lenslet images.

[0062] Furthermore, the size of the output virtual lenslet image is linearly larger than the s,t resolution of the input image, where the magnification n is adjustable depending on the settings.

[0063] FIG. 6 shows in more detail how a set of multiple lenslet images 10 is transformed into a virtual lenslet image.

[0064] 6, the lenslet synthesis processor 200 can grasp the positional relationship of the microlenses for each image frame by superimposing multiple image frames that make up the multiple lenslet image group 10. A virtual lenslet image is one in which virtual microlenses ML are arranged in image space, and the virtual microlenses ML may be overlapped in image space, or the virtual microlenses ML may be arranged in a non-overlapping manner within the virtual lenslet image according to their positional order.

[0065]

[0066] The sub-aperture image converter 300 can convert the virtual lenslet image into a sub-aperture image 30 .

[0067] At this time, the sub-aperture image conversion unit 300 can search for pixels corresponding to the coordinates of the microlens ML from the virtual lenslet image and map them to the coordinate position of the sub-aperture image to be converted.

[0068] FIG. 8 shows the processing steps of the sub-aperture image conversion unit 300.

[0069] As shown in Figure 8, the lenslet image contains information about the light passing through the arrangement of microlenses ML, resulting in a circular image appearing in the form of the arrangement. Depending on the number of microlenses ML in the image, the horizontal and vertical positions of each microlens ML are represented by s and t coordinates, and the number of microlenses ML in the image can be represented by the spatial resolution of the light field image. Each microlens ML has a predetermined number of pixel areas, and the position of each pixel within the microlens ML is represented by the u and v coordinates of the microlens ML, with the number of u and v representing the angular resolution of the light field image.

[0070] Since the position of each pixel in a light field image is aligned with s, t, u, and v coordinates, a process of converting the lenslet image into an aligned image is required, and the subaperture image conversion unit 300 performs a process of converting the lenslet image into a subaperture image. The process involves searching for pixels corresponding to s, t, u, and v coordinate positions in each lenslet image and mapping them to the s, t, v, and u image positions of the subaperture image. The converted subaperture image is composed of u×v block images having a size of s×t, and each block image has angle information corresponding to the u and v coordinates.

[0071] FIG. 7 shows an example of a method for generating pixel values ​​in a virtual lenslet image in the sub-aperture image conversion unit 300.

[0072] As shown in Figure 7, a virtual lenslet image Any lenslet image corresponding to positions i and j in TIFF2025508212000013.tif1216 has pixel sizes of u and v. To generate the lenslet image, a virtual lenslet image Adjacent lenslet images are found in TIFF2025508212000014.tif1216, and the pixel values ​​of the virtual lenslet image can be calculated using Equation 1 below.

[0073]

number

[0074]

[0075] where: TIFF2025508212000016.tif2774, TIFF2025508212000017.tif1449, JPEG2025508212000018.jpg1554 is the pixel value corresponding to the i, j, u, and v coordinates, TIFF2025508212000019.tif2025 is a weight value according to the distance between the kth lenslet image and the virtual lenslet image, TIFF2025508212000020.tif1619 is the distance between the kth lenslet image and the virtual lenslet image.

[0076]

[0077] As described above, the three-dimensional conversion unit 400 can convert the sub-aperture image generated by the sub-aperture image conversion unit 300 into a focal stack image.

[0078] FIG. 9 shows the processing steps of the three-dimensional conversion unit 400 that receives the sub-aperture image as input.

[0079] As shown in FIG. 9, the three-dimensional conversion unit 400 can convert the sub-aperture image into a focal stack image having three-dimensional information on the x-axis, y-axis, and z-axis.

[0080] The x-axis and y-axis positions of the focal stack image contain 2D image information at the corresponding z-axis coordinate point, and the depth information of the image can be determined according to the number of z-axis coordinates.

[0081] The three-dimensional conversion unit 400 can convert the sub-aperture image into a focal stack image using the following Equations 2 and 3.

[0082]

number

[0083]

[0084]

number

[0085]

[0086] where: JPEG2025508212000023.jpg2048 is the s, t, u, v coordinates and the transformed sub-aperture values ​​corresponding to a specific α value. The transformed sub-aperture values ​​are necessary to obtain a focal stack image, and the focal stack coordinate values ​​corresponding to the α value are JPEG2025508212000024.jpg1942 can be obtained. Note that x and y are x and y coordinate values ​​corresponding to the three-dimensional focal stack image. JPEG2025508212000025.jpg1115 is the number of u and v coordinates, where a specific z coordinate is matched to a specific α value, where α=1 corresponds to the center position of the focal cosine stack image, and has a range of 0<α<1+i, and the measurement range of the z axis can be adjusted by adjusting the range of α.

[0087]

[0088] Next, with reference to FIG. 11, a light field microscope-based image acquisition method according to an embodiment of the present invention will be described.

[0089] As shown in FIG. 11 , a light field microscope-based image acquisition method according to an embodiment of the present invention includes:

[0090] The method may include a first step S100 of acquiring a group of multiple lenslet images, a second step S200 of acquiring a virtual lenslet image using the group of multiple lenslet images acquired in the first step S100, a third step S300 of converting the virtual lenslet image acquired in the second step S200 into a subaperture image, and a fourth step S400 of converting the aperture image converted in the third step S300 into a focal stack image.

[0091] In the first step S100, multiple image frame information can be obtained through microlenses ML whose arrangement position is changed according to time period within a set image sensor area, and multiple lenslet image groups can be obtained by rotating a disk D on which multiple microlenses ML are arranged for a set time.

[0092] In the second step S200, the arrangement of microlenses for each image can be grasped by overlapping a plurality of image frames that constitute a multiple lenslet image group, and the second step S200 can be performed by the lenslet synthesis process 200.

[0093] In the third step S300, pixels corresponding to the coordinates of the microlenses in the virtual lenslet image can be searched for and mapped to the coordinate positions of the sub-aperture image to be converted, and the third step S300 can be performed by the sub-aperture image conversion unit 300.

[0094] In the fourth step S400, the sub-aperture image is converted into a focal stack image having three-dimensional information of the x-axis, y-axis, and z-axis, and the x-axis and y-axis positions of the focal stack image may include two-dimensional image information at the corresponding z-axis coordinate point. The fourth step S400 may be performed by the three-dimensional conversion unit 400.

[0095] The light field microscope-based image acquisition method according to one embodiment of the present invention generates high-resolution three-dimensional images based on the configuration described for the light field microscope-based image acquisition device 100 according to one embodiment of the present invention, and therefore, in order to avoid duplication of explanation, a detailed description thereof will be omitted.

[0096] Although specific embodiments of the present invention have been described above, it goes without saying that various modifications are possible without departing from the scope of the present invention.

[0097] Therefore, the scope of the present invention should not be limited to the above-described embodiments, but should be determined not only by the claims described below, but also by equivalents to the claims. [Industrial Applicability]

[0098] The present invention can be applied and sold in various industrial fields that use safety equipment, component inspection, material inspection, measurement and analysis instruments.

Claims

1. a lenslet synthesis processor that acquires a virtual lenslet image using a group of multiple lenslet images; a sub-aperture image conversion unit that converts the virtual lenslet image generated by the lenslet synthesis processing unit into a sub-aperture image; A light field microscope-based image acquisition device comprising: a three-dimensional conversion unit that converts the subaperture image generated by the subaperture image conversion unit into a focal stack image.

2. The multiple lenslet images include: The light field microscope-based image acquisition device of claim 1, wherein the image frame information is generated by acquiring multiple image frame information through microlenses whose arrangement positions are changed according to time periods within a set image sensor area.

3. The multiple lenslet images include: The light field microscope-based image acquisition device according to claim 2 , wherein the image is acquired by rotating a disk on which a plurality of microlenses are arranged for a set period of time.

4. The lenslet synthesis processing unit includes:

3. The light field microscope-based image acquisition device according to claim 2, wherein a plurality of image frames constituting a multi-lenslet image group are superimposed to grasp the arrangement relationship of the microlenses for each image.

5. The sub-aperture image conversion unit includes: The light field microscope-based image acquisition device according to claim 4 , characterized in that a pixel corresponding to the coordinates of the microlens in the virtual lenslet image is searched for and mapped to the coordinate position of the subaperture image to be converted.

6. The sub-aperture image conversion unit includes: The pixel values ​​of the virtual lenslet image are generated using the following Equation 1: [0010] Where: 【number】 and 【number】 and 【number】 are pixel values ​​corresponding to the i, j, u, and v coordinates, 【number】 is a weighting value according to the distance between the kth lenslet image and the virtual lenslet image, 【number】 The light field microscope based image acquisition device of claim 5 , wherein k is the distance between the kth lenslet image and the virtual lenslet image.

7. The three-dimensional conversion unit is Converting the subaperture image into a focal stack image having three-dimensional information of x-axis, y-axis, and z-axis; The light field microscope-based image acquisition device of claim 5 , wherein the x-axis and y-axis positions of the focal stack image include two-dimensional image information at a corresponding z-axis coordinate point.

8. The three-dimensional conversion unit converts the subaperture image into a focal stack image using the following Equations 2 and 3: [0025] [0030] Where: 【number】 are the transformed subaperture values ​​corresponding to s, t, u, v coordinates and a particular α value, 【number】 are focal stack coordinate values ​​corresponding to the α value, x, y are x, y coordinate values ​​corresponding to the three-dimensional focal stack image, 【number】 The light field microscope based image acquisition device of claim 7 , wherein x is the number of u,v coordinates.

9. The light field microscope based image acquisition device of claim 3 , wherein the disk is coupled to an actuator or motor that provides rotational or linear power.

10. The light field microscope-based image acquisition device of claim 3, wherein the linear movement range of the disk is 0-500 mm.

11. A first step of acquiring multiple lenslet images; a second step of acquiring a virtual lenslet image using the multiple lenslet images acquired in the first step; a third step of converting the virtual lenslet image acquired in the second step into a subaperture image; and a fourth step of converting the subaperture images converted in the third step into focal stack images.

12. In the first stage, The light field microscope-based image acquisition method according to claim 11, further comprising acquiring a plurality of image frame information through a microlens whose arrangement position is changed according to time period within a set image sensor area.

13. In the first stage, The light field microscope-based image acquisition method according to claim 12, characterized in that a disk on which a plurality of microlenses are arranged is rotated for a set period of time to acquire a group of multiple lenslet images.

14. The light field microscope-based image acquisition method of claim 12, wherein in the second step, a plurality of image frames constituting a group of multiple lenslet images are superimposed to grasp the arrangement relationship of microlenses for each image.

15. 15. The light field microscope-based image acquisition method of claim 14, wherein in the third step, a pixel corresponding to the coordinate of the microlens is searched for in the virtual lenslet image and mapped to the coordinate position of the subaperture image to be converted.

16. In the third step, pixel values ​​of the virtual lenslet image are generated using the following Equation 1: [0010] Where: 【number】 and 【number】 and 【number】 are pixel values ​​corresponding to the i, j, u, and v coordinates, 【number】 is a weighting value according to the distance between the kth lenslet image and the virtual lenslet image, 【number】 The light field microscope based image acquisition method of claim 15, wherein k is the distance between the kth lenslet image and the virtual lenslet image.

17. In the fourth stage, Converting the subaperture image into a focal stack image having three-dimensional information of x-axis, y-axis, and z-axis; The light field microscope-based image acquisition method of claim 15, wherein the x-axis and y-axis positions of the focal stack image include two-dimensional image information at a corresponding z-axis coordinate point.

18. In the fourth step, the sub-aperture image is converted into a focal stack image using the following Equations 2 and 3: [0025] [0030] Where: 【number】 are the transformed subaperture values ​​corresponding to s, t, u, v coordinates and a particular α value, 【number】 are focal stack coordinate values ​​corresponding to the α value, x, y are x, y coordinate values ​​corresponding to the three-dimensional focal stack image, 【number】 The light field microscopy based image acquisition method of claim 17, wherein x is the number of u,v coordinates.

Images