Cell sample imaging method, system and equipment and storage medium

Abstract

The invention discloses a cell sample imaging method, system and device and a storage medium, and the method comprises the steps: determining an initial focus parameter of a three-dimensional control platform for collecting a transparent carrier, determining a collection focus height when traversing to a preset laser position of each collection unit of each bearing position, and determining a collection focus parameter of each collection unit of each bearing position; in the process that the horizontal motion platform moves from the preset laser position to the sampling position, the vertical shaft motor synchronously moves to the collection focus height of the collection unit of the bearing position, and the cell image of the bearing position at the sampling position of the collection unit is collected according to the collection focus height of the collection unit of the bearing position. Therefore, the bright field scanning and the laser ranging are carried out at the same time, and in the bright field scanning process, the vertical shaft motor moves synchronously, so that the waiting time in the bright field scanning process is shortened, and the bright field scanning efficiency in the cell imaging process is improved.

Description

Technical Field

[0001] This application relates to the field of biological image acquisition, and more specifically, to a method, system, device, and storage medium for cell sample imaging. Background Technology

[0002] With the rapid development of cell technology, cell imaging is being used more and more widely in fields such as biology and chemistry. As a core optical instrument, the microscope magnifies and images tiny cell samples through the objective lens, providing key support for operators to observe cell details.

[0003] In modern cell imaging systems, bright-field scanning is a crucial step in acquiring images of samples on transparent carriers. The core workflow of bright-field scanning is as follows: the XY platform moves to the target imaging position and remains stationary, while the Z-axis motor moves from the preset position Z(X,Y) to two positions, Z(X,Y)+DF and Z(X,Y)-DF. After each movement, the camera is triggered to capture and save the image. After completing one column of scanning, the XY platform moves to the next column and repeats the operation.

[0004] However, the Z-axis motor needs to reciprocate between two fixed positions, and the XY platform must remain stationary during image acquisition, resulting in long waiting times and cumbersome operation steps for the entire bright field scanning process, which severely restricts scanning efficiency.

[0005] Therefore, improving the efficiency of bright-field scanning during cell imaging is an issue that needs attention. Summary of the Invention

[0006] In view of the above problems, this application provides a cell sample imaging method, system, device and storage medium to improve the efficiency of bright field scanning in the cell imaging process.

[0007] To achieve the above objectives, the following specific solutions are proposed:

[0008] A cell sample imaging method is applied to a cell sample imaging system, the cell sample imaging system comprising a three-dimensional control platform, a bright field light source, a laser light source, an objective lens, a transparent carrier, and an imaging camera;

[0009] The three-dimensional control platform includes a horizontal motion platform and a vertical axis motor. The vertical axis motor is fixed with the objective lens. The transparent carrier is placed on the horizontal motion platform. The transparent carrier contains multiple support positions for culturing cells. The bright field light source emits a beam that passes through the transparent carrier and the objective lens in sequence and is then received by the imaging camera. The laser light source is located between the objective lens and the transparent carrier.

[0010] The method includes:

[0011] Determine the initial focal parameters for the three-dimensional control platform to collect data from the transparent carrier;

[0012] In the process of taking bright-field photographs by traversing each carrier position of the transparent carrier using the initial focal parameters as the initial parameters for bright-field photographing, when the preset laser position of each acquisition unit of each carrier position is reached, the acquisition focal height of that carrier position for bright-field photographing in that acquisition unit is determined.

[0013] During the process of the horizontal motion platform moving from the preset laser position of each acquisition unit of each bearing position to the sampling position of that acquisition unit of that bearing position, the vertical axis motor is driven to move to the acquisition focal height of that acquisition unit of that bearing position;

[0014] The cell image at the sampling position of each acquisition unit is acquired using the acquisition focal height of each acquisition unit at each acquisition position.

[0015] Optionally, the imaging camera is located vertically below the objective lens;

[0016] Determining the initial focal parameters for the 3D control platform to acquire data from the transparent carrier includes:

[0017] The initial bearing position of the transparent carrier is moved above the objective lens by the horizontal motion platform.

[0018] With both the bright field light source and the imaging camera turned on, the vertical axis motor is adjusted and multiple images of the transparent carrier surface at different focus heights are acquired. The focus height corresponding to the transparent carrier surface image with the highest clarity among the transparent carrier surface images is determined as the initial focal plane height.

[0019] When the vertical axis motor is at the initial focal plane height, the laser source is turned on, the center position of the laser spot in the light spot image acquired by the imaging camera is recorded, and the initial focal parameters are obtained by combining the initial focal plane height and the center position of the laser spot.

[0020] Optionally, when traversing to the preset laser position of each acquisition unit of each carrier position, determining the acquisition focal height of that carrier position for bright-field photography in that acquisition unit includes:

[0021] For each acquisition unit of each traversed bearing position, a laser image is acquired at the preset laser position with the laser source turned on, and the center position of the light spot in the laser image is determined.

[0022] For each acquisition unit at each traversed carrier position, the center position of the laser spot at the preset laser position is calculated, and the deviation value between the center position of the laser spot and the latest laser spot center position is calculated. Based on the deviation value, the acquisition focal height of the acquisition unit at the carrier position for bright-field photography is calculated.

[0023] Optionally, each carrier position in the transparent carrier is arranged in a row and column matrix, each acquisition unit of each carrier position is arranged in a row and column matrix, and all acquisition units in the transparent carrier are arranged in a row and column matrix.

[0024] The method also includes:

[0025] The horizontal motion platform is driven to move to the initial acquisition unit of the transparent carrier. When the horizontal motion platform moves to each acquisition unit, the cell imaging task of that acquisition unit is executed.

[0026] After completing the cell imaging task of each acquisition unit, the horizontal motion platform is driven to move in the first horizontal direction until it reaches the next acquisition unit and performs its cell imaging task.

[0027] After the horizontal motion platform moves to the last acquisition unit in the first horizontal direction and completes its cell imaging task, it is driven to move to the second horizontal direction to the adjacent acquisition unit that has not yet been imaged and to perform its cell imaging task. Then, the horizontal motion platform is driven to move in the opposite direction of the current first horizontal direction to continue to image the acquisition units in the current first horizontal direction one by one.

[0028] After the horizontal motion platform moves to the last acquisition unit in the second horizontal direction, the cell imaging task of the transparent carrier ends when the cell imaging task of the last acquisition unit in the current first horizontal dimension is completed.

[0029] A cell sample imaging system includes a three-dimensional control platform, a bright field light source, a laser light source, an objective lens, a transparent carrier, an imaging camera, and a controller;

[0030] The three-dimensional control platform includes a horizontal motion platform and a vertical axis motor. The objective lens is fixed to the vertical axis motor. The transparent carrier is placed on the horizontal motion platform. The transparent carrier contains multiple support positions for culturing cells. The bright field light source, the objective lens, the transparent carrier, and the laser light source are coaxial from top to bottom in the vertical direction.

[0031] The controller includes:

[0032] A focal parameter initialization unit is used to determine the initial focal parameters for the three-dimensional control platform to collect data from the transparent carrier.

[0033] The focal height determination unit is used to determine the focal height of the carrier for bright-field photography when, during the process of traversing each carrier position of the transparent carrier to perform bright-field photography using the initial focal parameters as the initial parameters for bright-field photography traversal, the carrier position is traversed to the preset laser position of each acquisition unit of each carrier position.

[0034] A synchronous moving unit is used to drive the vertical axis motor to move to the acquisition focal height of the acquisition unit at the bearing position during the process of the horizontal motion platform moving from the preset laser position of each acquisition unit at each bearing position to the sampling position of the acquisition unit at that bearing position;

[0035] An imaging unit is used to acquire a cell image of the carrier at the sampling position of the acquisition unit at the acquisition focal height of each acquisition unit of each carrier position.

[0036] Optionally, the imaging camera is located vertically below the objective lens;

[0037] The focal parameter initialization unit includes:

[0038] The first focal parameter initialization subunit is used to move the initial bearing position of the transparent carrier to above the objective lens via the horizontal motion platform.

[0039] The second focal parameter initialization subunit is used to adjust the vertical axis motor and acquire multiple transparent carrier surface images with different focus heights when both the bright field light source and the imaging camera are turned on, and to determine the focus height corresponding to the transparent carrier surface image with the highest clarity among the transparent carrier surface images as the initial focal plane height.

[0040] The third focal parameter initialization subunit is used to turn on the laser source when the vertical axis motor is at the initial focal plane height, record the center position of the laser spot in the light spot image acquired by the imaging camera, and combine the initial focal plane height and the center position of the laser spot to obtain the initial focal parameters.

[0041] Optionally, the focal height determination unit includes:

[0042] The spot center determination unit is used to acquire a laser image at the preset laser position for each acquisition unit of each traversed bearing position under the condition that the laser source is turned on, and to determine the center position of the spot in the laser image.

[0043] The focal height calculation unit is used to calculate the deviation between the center position of the laser spot and the latest center position of the laser spot for each acquisition unit at each traversed carrier position, and to calculate the focal height of the acquisition unit at the carrier position for bright-field photography based on the deviation value.

[0044] Optionally, each carrier position in the transparent carrier is arranged in a row and column matrix, each acquisition unit of each carrier position is arranged in a row and column matrix, and all acquisition units in the transparent carrier are arranged in a row and column matrix.

[0045] The controller also includes:

[0046] The acquisition unit task execution unit is used to drive the horizontal motion platform to move to the initial acquisition unit of the transparent carrier. When the horizontal motion platform moves to each acquisition unit, it executes the cell imaging task of that acquisition unit.

[0047] A one-dimensional horizontal movement unit is used to drive the horizontal motion platform to move in the first-dimensional horizontal direction after completing the cell imaging task of each acquisition unit, until it reaches the next acquisition unit of that acquisition unit and performs its cell imaging task.

[0048] A two-dimensional horizontal movement unit is used to drive the horizontal motion platform to move in the second dimension to the adjacent acquisition unit that has not yet been imaged and to perform its cell imaging task after the horizontal motion platform moves in the first dimension horizontal direction to the last acquisition unit and completes its cell imaging task. It also drives the horizontal motion platform to move in the opposite direction of the current first dimension horizontal direction to continue to image the acquisition units in the current first dimension horizontal direction one by one.

[0049] The imaging task completion unit is used to end the cell imaging task of the transparent carrier after the horizontal motion platform moves to the last acquisition unit in the second horizontal direction and completes the cell imaging task of the last acquisition unit in the current first horizontal dimension.

[0050] A cell sample imaging device, including a memory and a processor;

[0051] The memory is used to store programs;

[0052] The processor is used to execute the program to implement the various steps of the cell sample imaging method described above.

[0053] A storage medium having a computer program stored thereon, which, when executed by a processor, implements the various steps of the cell sample imaging method as described above.

[0054] By employing the above technical solution, this application determines the initial focal parameters for the 3D control platform to acquire data from the transparent carrier. Using these initial focal parameters as the initial parameters for bright-field imaging traversal, the platform traverses each carrier position of the transparent carrier for bright-field imaging. When traversing to the preset laser position of each acquisition unit at each carrier position, the acquisition focal height for that carrier position at that acquisition unit is determined. As the horizontal motion platform moves from the preset laser position of each acquisition unit at each carrier position to the sampling position of that acquisition unit, the vertical axis motor is driven to move to the acquisition focal height of that acquisition unit at each carrier position. Cell images at the sampling position of that carrier position at each acquisition unit are acquired using the acquisition focal height of that acquisition unit. Therefore, bright-field scanning and laser ranging are performed simultaneously. During bright-field scanning, the vertical axis motor moves synchronously, reducing the waiting time during bright-field scanning and improving the efficiency of bright-field scanning in cell imaging. Attached Figure Description

[0055] Various other advantages and benefits will become apparent to those skilled in the art upon reading the following detailed description of preferred embodiments. The accompanying drawings are for illustrative purposes only and are not intended to limit the scope of this application. Furthermore, the same reference numerals denote the same parts throughout the drawings. In the drawings:

[0056] Figure 1 This is a schematic diagram of a system structure for realizing cell sample imaging provided in an embodiment of this application;

[0057] Figure 2 This is a schematic diagram of a process for achieving cell sample imaging provided in an embodiment of this application;

[0058] Figure 3 A layout diagram of a transparent carrier provided in an embodiment of this application;

[0059] Figure 4 A schematic diagram of a data acquisition unit for a transparent carrier bearing position provided in an embodiment of this application;

[0060] Figure 5 A schematic diagram of a process for obtaining initial focal parameters provided in an embodiment of this application;

[0061] Figure 6 A schematic diagram of a transparent carrier bright-field traversal cell imaging process provided for an embodiment of this application;

[0062] Figure 7 This application provides a motor signal triggering diagram for full-panel bright-field imaging of a transparent carrier.

[0063] Figure 8This application provides a schematic diagram of the unit structure of a cell sample imaging system processor for implementing cell sample imaging.

[0064] Figure 9 This is a schematic diagram of the structure of a device for imaging cell samples provided in an embodiment of this application. Detailed Implementation

[0065] The technical solutions of the embodiments of this application will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of this application, and not all embodiments. Based on the embodiments of this application, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of this application.

[0066] This application provides an optional system architecture for imaging cell samples. This system architecture may include a three-dimensional control platform, a bright field light source, a laser light source, an objective lens, a transparent carrier, an imaging camera, a memory, and a controller.

[0067] The three-dimensional control platform can include a horizontal motion platform (XY motion platform), a horizontal axis motor (X motor), a vertical axis motor (Y motor), and a vertical axis motor (Z motor). The Z motor is fixed with an objective lens. A transparent carrier is placed on the horizontal motion platform. The transparent carrier contains multiple carrier sites for cell culture. These carrier sites can be arranged in a row-column array. Different types of transparent carriers have different numbers of carrier sites, and the size of each carrier site also varies. For example, in a transparent carrier with 96 (12×8) carrier sites and a transparent carrier with 384 (24×16) carrier sites, the carrier site size of the 96-well plate is larger than that of the 384-well plate. The imaging camera is located vertically below the laser source. The bright-field light source emits a beam that passes sequentially through the transparent carrier and the objective lens before being received by the imaging camera. The laser source is located between the objective lens and the transparent carrier, and the imaging camera is located vertically below the objective lens. Each carrier site can contain multiple acquisition units, and these acquisition units can be arranged in a row-column array. Therefore, all acquisition units within the transparent carrier can be arranged in a row-column array. Each acquisition unit can contain a preset laser position and a sampling position. The system can first focus at the preset laser position and then move to the sampling position to acquire the image.

[0068] Specifically, the X motor has its corresponding X motor encoder, the Y motor has its corresponding Y motor encoder, and the Z motor has its corresponding Z motor encoder. The signal flow and control method of this system can be found in [reference needed]. Figure 1The controller can send corresponding motion commands to the laser light source, bright field light source, imaging camera, memory, and XYZ motors. The controller can also acquire position data stored in the memory and encoder data representing the movement of the XYZ motors. The XY motors are used to control the movement of the XY motion platform in the horizontal two-dimensional direction, while the Z motors are used to control the movement of the transparent carrier in the Z-axis direction.

[0069] Based on the above system architecture, system signal flow, and control method, Figure 2 This illustration shows a flowchart of a cell sample imaging method provided in an embodiment of this application. (Refer to...) Figure 2 The process may include:

[0070] Step S110: Determine the initial focal parameters for the 3D control platform to collect data from the transparent carrier.

[0071] Understandably, the initial focal plane height of the 3D control platform is determined before bright-field scanning acquisition. This determines the initial focal plane at that height, providing an approximate position for focusing the carrier / acquisition unit. This facilitates subsequent focus measurement of the carrier / acquisition unit and avoids significant adjustments to the Z motor during focus measurement at the carrier position.

[0072] Step S120: During the process of traversing each carrier position of the transparent carrier to perform bright-field photography using the initial focal parameters as the initial parameters for bright-field photography traversal, when the preset laser position of each acquisition unit of each carrier position is reached, the acquisition focal height of that carrier position for bright-field photography in that acquisition unit is determined.

[0073] For example Figure 3 and Figure 4 For a transparent carrier with 96 (12×8) carrier sites, each carrier site can contain 4×4 acquisition units. Therefore, each carrier site contains 16 preset laser positions and corresponding sampling positions. After determining the acquisition focal height at the preset laser positions, cell images can be acquired at the sampling positions according to the acquisition focal height.

[0074] Step S130: During the process of the horizontal motion platform moving from the preset laser position of each acquisition unit of each bearing position to the sampling position of the acquisition unit of that bearing position, the vertical axis motor is driven to move to the acquisition focal height of the acquisition unit of that bearing position.

[0075] Understandably, while the XY motors move, the Z motor does not need to remain stationary and wait for the XY motors to complete their adjustments before moving. The Z motor and XY motors can move synchronously, thereby reducing the waiting time during bright-field scanning and improving the efficiency of bright-field scanning in cell imaging.

[0076] It should be noted that since the XY motor will trigger the shooting immediately after moving to the sampling position, the Z motor needs to be adjusted to move the sampling focus height before the XY motor moves to the sampling position.

[0077] Step S140: Acquire the cell image at the sampling position of each acquisition unit of each carrier position using the acquisition focal height of each acquisition unit.

[0078] Specifically, after the Z motor is adjusted to the acquisition focal height and the XY motor is moved to the sampling position, the actions of turning on the bright field light source → turning on the imaging camera → acquiring bright field cell images → turning off the bright field are executed in sequence, thereby completing cell imaging at the sampling position.

[0079] The cell sample imaging method provided in this embodiment determines the initial focal parameters for the three-dimensional control platform to acquire data from a transparent carrier. Using these initial focal parameters as the initial parameters for bright-field imaging traversal, the method traverses each carrier position on the transparent carrier for bright-field imaging. When the traversal reaches the preset laser position of each acquisition unit at each carrier position, the acquisition focal height for that carrier position at that acquisition unit is determined. As the horizontal motion platform moves from the preset laser position of each acquisition unit at each carrier position to the sampling position of that acquisition unit, the vertical axis motor is driven to move to the acquisition focal height of that acquisition unit at each carrier position. Cell images at the sampling position of that carrier position at each acquisition unit are acquired using the acquisition focal height of that acquisition unit. Therefore, bright-field scanning and laser ranging are performed simultaneously. During bright-field scanning, the vertical axis motor moves synchronously, reducing the waiting time during bright-field scanning and improving the efficiency of bright-field scanning in cell imaging.

[0080] In some embodiments of this application, the process of determining the initial focal parameters for the three-dimensional control platform to acquire data from the transparent carrier in step S110 is described. This process may include:

[0081] S1. Move the initial bearing position of the transparent carrier to above the objective lens using a horizontal motion platform.

[0082] S2. With both the bright field light source and the imaging camera on, adjust the vertical axis motor and acquire multiple images of the transparent carrier surface at different focus heights. Determine the focus height corresponding to the transparent carrier surface image with the highest clarity among the images as the initial focal plane height.

[0083] S3. With the vertical axis motor at the initial focal plane height, turn on the laser source, record the center position of the laser spot in the image of the light spot acquired by the imaging camera, and combine the initial focal plane height and the center position of the laser spot to obtain the initial focal parameters.

[0084] For example Figure 5 The initial focus parameter acquisition process is as follows: the controller controls the XY motion platform to move to the A1 bearing position of the transparent carrier (e.g., Figure 3 (The A1 bearing position shown is the initial bearing position); whenever the Z motor stops moving, the bright field light source and imaging camera are turned on; adjust Z until a clear image of the transparent carrier surface is acquired and the position is recorded as the initial focal plane height Z0. Specifically, the Z motor can be moved from the low point Zori to the end point, i.e., the highest point Ztar. The Z motor movement step size can be set as Zgap, and the number of Z motor step movements can be set as n. Therefore, the movement function of the Z motor is Ztar = Zori + Zgap * n. While moving, images of the transparent carrier surface are acquired, and finally n images of the transparent carrier surface are acquired. Finally, the image with the best clarity is identified from multiple images of the transparent carrier surface, and the corresponding focus height is determined as the initial focal plane height; then the bright field light source is turned off. Further, the laser light source is turned on again at the initial focal plane height Z0, and the imaging camera can acquire a spot image; record the XY position (X0, Y0) of the center of the spot at this time, i.e., the center position of the laser spot; then the laser light source is turned off. By combining the XY plane position (X0, Y0) and the initial focal plane height Z0, the initial focal parameters can be obtained as (X0, Y0, Z0).

[0085] In some embodiments of this application, the process of determining the focal height for bright-field photography of each acquisition unit when traversing to the preset laser position of each acquisition unit of each carrier position, as mentioned in the above embodiments, is described. This process may include:

[0086] S1. For each acquisition unit of each traversed bearing position, acquire the laser image at the preset laser position with the laser light source turned on, and determine the center position of the light spot in the laser image.

[0087] For example Figure 3 and Figure 4 When traversing to the first acquisition unit in the first column of the bearing position A1, the horizontal motion platform can be moved to position L1 of that acquisition unit, then the laser light source can be turned on, the camera can be turned on to acquire the laser image, and the laser can be turned off. At this time, the center position (X, Y) of the laser spot can be obtained.

[0088] S2. For the center position of the laser spot of each acquisition unit at each traversed carrier position, calculate the deviation value between the center position of the laser spot and the latest laser spot center position, and calculate the acquisition focal height of the acquisition unit at the carrier position for bright field photography based on the deviation value.

[0089] The current and latest laser spot center position of the initial acquisition unit is the laser spot center position during the initial focus parameter determination process. The current and latest laser spot center position of non-initial acquisition units is the spot center position of the preset laser position of the previous acquisition unit.

[0090] For example Figure 3 and Figure 4 When the center position (X, Y) of the laser image of the first acquisition unit in the first column of the carrier position A1 is obtained, the deviation value between the center position (X, Y) and the center position (X0, Y0) of the laser image determined during initialization can be calculated, and the required acquisition focal height of the acquisition unit when taking pictures in bright field can be calculated based on the deviation value.

[0091] Furthermore, when the horizontal motion platform continues to move along the Y direction to the sampling position C1 of the first acquisition unit in the first column of the bearing position A1, the bright field light source can be turned on, the camera can be turned on to acquire a bright field image, and then the bright field light source can be turned off to complete the cell imaging task of that acquisition unit.

[0092] Furthermore, after completing the cell imaging task of the first acquisition unit in the first column of carrier position A1, the center position (X, Y) of the laser image at the preset laser position can be used as the latest center position (X0, Y0). When it is necessary to calculate the acquisition focal height of the next acquisition unit (the second acquisition unit in the first column of A1), it can be calculated based on the deviation between the center position of the acquisition unit and the latest center position (X0, Y0).

[0093] In the following embodiments, the process of acquiring full-field images of a transparent carrier using a 3D control platform will be described. Please refer to [link / reference needed] for details. Figure 6 This process needs to follow the following elements:

[0094] First, the horizontal motion platform is driven to move to the initial acquisition unit of the transparent carrier. When the horizontal motion platform moves to each acquisition unit, the cell imaging task of that acquisition unit is executed.

[0095] For example, first drive the horizontal motion platform to move to, such as Figure 3 and Figure 4 The initial acquisition unit of the transparent carrier shown (the first acquisition unit in the first column of the initial carrier position A1) performs cell imaging tasks of L1 focusing and C1 imaging within the acquisition unit.

[0096] Then, after completing the cell imaging task for each acquisition unit, the horizontal motion platform is driven to move in the first horizontal direction until it reaches the next acquisition unit and performs its cell imaging task.

[0097] The first horizontal direction can be either the Y-direction or the X-direction of the horizontal motion platform.

[0098] For example Figure 3 and Figure 4 After completing the cell imaging task for the first acquisition unit in the first column of carrier position A1, the Y motor can continue to move along the Y direction until it reaches the second acquisition unit in the first column of the initial carrier position A1, and perform the cell imaging task for that acquisition unit (perform L2 focusing and C2 image acquisition). It should be noted that after traversing and completing the cell imaging task for the fourth acquisition unit in the first column of carrier position A1 (that is, the last acquisition unit of the carrier position in the first dimension), the Y motor needs to continue to move to the first acquisition unit in the first column of carrier position B1. Therefore, the distance between the fourth acquisition unit in the first column of carrier position A1 and the first acquisition unit in the first column of carrier position B1 can be pre-calculated, and the Y motor can be driven to move from the fourth acquisition unit in the first column of carrier position A1 to the first acquisition unit in the first column of carrier position B1 based on this distance.

[0099] Furthermore, after the horizontal motion platform moves to the last acquisition unit in the first horizontal direction and completes its cell imaging task, it is driven to move to the second horizontal direction to the adjacent acquisition unit that has not yet undergone cell imaging and to perform its cell imaging task. Then, the horizontal motion platform is driven to move in the opposite direction of the current first horizontal direction to continue to perform cell imaging on the acquisition units in the current first horizontal direction one by one.

[0100] The second horizontal direction is perpendicular to the first horizontal direction on the horizontal plane.

[0101] For example Figure 3 and Figure 4After the horizontal motion platform moves to the fourth acquisition unit in the first column of carrier position H1 (the last acquisition unit of the last carrier position in the first horizontal dimension) and completes its cell imaging task, the cell imaging task for the next column of acquisition units will begin. Specifically, the X motor of the horizontal motion platform can be driven to translate to the acquisition unit adjacent to the fourth acquisition unit in the first column of carrier position H1. At this time, the Y motor needs to move in the opposite direction in the first horizontal dimension, that is, move the first acquisition unit in the second column of carrier position H1. Then, cell imaging continues to traverse all acquisition units in the second column, that is, traverse from the first acquisition unit in the second column of carrier position H1 to the fourth acquisition unit in the second column of carrier position A1. It is important to note that after traversing and completing the cell imaging task of the 4th unit in the 4th column of carrier position A1, the X-motor needs to be moved to the 1st unit in the 1st column of carrier position A2. Therefore, the distance between the 4th unit in the 4th column of carrier position A1 and the 1st unit in the 1st column of carrier position A2 can be pre-calculated, and the X-motor can be driven to move from the 4th unit in the 4th column of carrier position A1 to the 1st unit in the 1st column of carrier position A2 based on this distance. Cell imaging of each acquisition unit on the transparent carrier is performed in this serpentine traversal manner.

[0102] Finally, after the horizontal motion platform moves to the last acquisition unit in the second horizontal direction, the cell imaging task of the transparent carrier ends when the cell imaging task of the last acquisition unit in the current first horizontal dimension is completed.

[0103] Understandably, once the cell imaging task of the last acquisition unit in the last column of the transparent carrier is completed, it indicates that all cell samples on the transparent carrier have been acquired, thus completing and ending the current cell imaging task on the transparent carrier. The timing signal of the 3D control platform during bright-field acquisition of the transparent carrier can be referenced... Figure 7 .

[0104] The controller for cell sample imaging provided in the embodiments of this application is described below. The controller for cell sample imaging described below can be referred to in correspondence with the method for cell sample imaging described above.

[0105] See Figure 8 , Figure 8 This is a schematic diagram of a unit structure for a controller to achieve cell sample imaging, as disclosed in an embodiment of this application.

[0106] like Figure 8 As shown, the controller may include:

[0107] The focal parameter initialization unit 100 is used to determine the initial focal parameters for the three-dimensional control platform to collect data from the transparent carrier.

[0108] The focal height determination unit 200 is used to determine the focal height of the carrier for bright-field photography when the carrier is traversed to each carrier position of the transparent carrier in the process of taking bright-field photographs using the initial focal parameters as the initial parameters for bright-field photography traversal.

[0109] The synchronous moving unit 300 is used to drive the vertical axis motor to move to the acquisition focal height of the acquisition unit at the bearing position during the process of the horizontal motion platform moving from the preset laser position of each acquisition unit at each bearing position to the sampling position of the acquisition unit at that bearing position;

[0110] Imaging unit 400 is used to acquire cell images of the carrier at the sampling position of the acquisition unit at the acquisition focal height of each acquisition unit of each carrier position.

[0111] Optionally, the focus parameter initialization unit includes:

[0112] The first focal parameter initialization subunit is used to move the initial bearing position of the transparent carrier to above the objective lens via the horizontal motion platform.

[0113] The second focal parameter initialization subunit is used to adjust the vertical axis motor and acquire multiple transparent carrier surface images with different focus heights when both the bright field light source and the imaging camera are turned on, and to determine the focus height corresponding to the transparent carrier surface image with the highest clarity among the transparent carrier surface images as the initial focal plane height.

[0114] The third focal parameter initialization subunit is used to turn on the laser source when the vertical axis motor is at the initial focal plane height, record the center position of the laser spot in the light spot image acquired by the imaging camera, and combine the initial focal plane height and the center position of the laser spot to obtain the initial focal parameters.

[0115] Optionally, the focal height determination unit includes:

[0116] The spot center determination unit is used to acquire a laser image at the preset laser position for each acquisition unit of each traversed bearing position under the condition that the laser source is turned on, and to determine the center position of the spot in the laser image.

[0117] The focal height calculation unit is used to calculate the deviation between the center position of the laser spot and the latest center position of the laser spot for each acquisition unit at each traversed carrier position, and to calculate the focal height of the acquisition unit at the carrier position for bright-field photography based on the deviation value.

[0118] Optionally, each carrier position in the transparent carrier is arranged in a row and column matrix, each acquisition unit of each carrier position is arranged in a row and column matrix, and all acquisition units in the transparent carrier are arranged in a row and column matrix.

[0119] The controller also includes:

[0120] The acquisition unit task execution unit is used to drive the horizontal motion platform to move to the initial acquisition unit of the transparent carrier. When the horizontal motion platform moves to each acquisition unit, it executes the cell imaging task of that acquisition unit.

[0121] A one-dimensional horizontal movement unit is used to drive the horizontal motion platform to move in the first-dimensional horizontal direction after completing the cell imaging task of each acquisition unit, until it reaches the next acquisition unit of that acquisition unit and performs its cell imaging task.

[0122] A two-dimensional horizontal movement unit is used to drive the horizontal motion platform to move in the second dimension to the adjacent acquisition unit that has not yet been imaged and to perform its cell imaging task after the horizontal motion platform moves in the first dimension horizontal direction to the last acquisition unit and completes its cell imaging task. It also drives the horizontal motion platform to move in the opposite direction of the current first dimension horizontal direction to continue to image the acquisition units in the current first dimension horizontal direction one by one.

[0123] The imaging task completion unit is used to end the cell imaging task of the transparent carrier after the horizontal motion platform moves to the last acquisition unit in the second horizontal direction and completes the cell imaging task of the last acquisition unit in the current first horizontal dimension.

[0124] The cell sample imaging method provided in this application embodiment can be applied to cell sample imaging equipment. Optionally, Figure 9 A hardware block diagram of the cell sample imaging device is shown, with reference to... Figure 9 The hardware structure of a cell sample imaging device may include: at least one processor 1, at least one communication interface 2, at least one memory 3, and at least one communication bus 4;

[0125] In this embodiment of the application, the number of processor 1, communication interface 2, memory 3, and communication bus 4 is at least one, and processor 1, communication interface 2, and memory 3 communicate with each other through communication bus 4;

[0126] Processor 1 may be a central processing unit (CPU), an application-specific integrated circuit (ASIC), a field-programmable gate array (FPGA), or one or more integrated circuits configured to implement embodiments of the present invention.

[0127] Memory 3 may include high-speed RAM, and may also include non-volatile memory, such as at least one disk storage device;

[0128] The memory stores a program, which the processor can call. The program is used for:

[0129] Determine the initial focal parameters for the 3D control platform to acquire data from the transparent carrier;

[0130] In the process of taking bright-field photographs by traversing each carrier position of the transparent carrier using the initial focal parameters as the initial parameters for bright-field photographing, when the preset laser position of each acquisition unit of each carrier position is reached, the acquisition focal height of that carrier position for bright-field photographing in that acquisition unit is determined.

[0131] During the process of the horizontal motion platform moving from the preset laser position of each acquisition unit of each bearing position to the sampling position of that acquisition unit of that bearing position, the vertical axis motor is driven to move to the acquisition focal height of that acquisition unit of that bearing position;

[0132] The cell image at the sampling position of each acquisition unit is acquired using the acquisition focal height of each acquisition unit at each acquisition position.

[0133] Optionally, the refined and extended functions of the program can be found in the description above.

[0134] This application embodiment also provides a storage medium that can store a program suitable for execution by a processor, the program being used for:

[0135] Determine the initial focal parameters for the 3D control platform to acquire data from the transparent carrier;

[0136] In the process of taking bright-field photographs by traversing each carrier position of the transparent carrier using the initial focal parameters as the initial parameters for bright-field photographing, when the preset laser position of each acquisition unit of each carrier position is reached, the acquisition focal height of that carrier position for bright-field photographing in that acquisition unit is determined.

[0137] During the process of the horizontal motion platform moving from the preset laser position of each acquisition unit of each bearing position to the sampling position of that acquisition unit of that bearing position, the vertical axis motor is driven to move to the acquisition focal height of that acquisition unit of that bearing position;

[0138] The cell image at the sampling position of each acquisition unit is acquired using the acquisition focal height of each acquisition unit at each acquisition position.

[0139] Optionally, the refined and extended functions of the program can be found in the description above.

[0140] Finally, it should be noted that in this document, relational terms such as "first" and "second" are used only to distinguish one entity or operation from another, and do not necessarily require or imply any such actual relationship or order between these entities or operations. Furthermore, the terms "comprising," "including," or any other variations thereof are intended to cover non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements includes not only those elements but also other elements not expressly listed, or elements inherent to such a process, method, article, or apparatus. Without further limitations, an element defined by the phrase "comprising one..." does not exclude the presence of other identical elements in the process, method, article, or apparatus that includes said element.

[0141] The various embodiments in this specification are described in a progressive manner. Each embodiment focuses on the differences from other embodiments. The various embodiments can be combined as needed, and the same or similar parts can be referred to each other.

[0142] The above description of the disclosed embodiments enables those skilled in the art to make or use this application. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of this application. Therefore, this application is not to be limited to the embodiments shown herein, but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. A method for imaging cell samples, characterized in that, An application in a cell sample imaging system, the cell sample imaging system comprising a three-dimensional control platform, a bright field light source, a laser light source, an objective lens, a transparent carrier, and an imaging camera; The three-dimensional control platform includes a horizontal motion platform and a vertical axis motor. The vertical axis motor is fixed with the objective lens. The transparent carrier is placed on the horizontal motion platform. The transparent carrier contains multiple support positions for culturing cells. The bright field light source emits a beam that passes through the transparent carrier and the objective lens in sequence and is then received by the imaging camera. The laser light source is located between the objective lens and the transparent carrier. The method includes: Determine the initial focal parameters for the three-dimensional control platform to collect data from the transparent carrier; In the process of taking bright-field photographs by traversing each carrier position of the transparent carrier using the initial focal parameters as the initial parameters for bright-field photographing, when the preset laser position of each acquisition unit of each carrier position is reached, the acquisition focal height of that carrier position for bright-field photographing in that acquisition unit is determined. During the process of the horizontal motion platform moving from the preset laser position of each acquisition unit of each bearing position to the sampling position of that acquisition unit of that bearing position, the vertical axis motor is driven to move to the acquisition focal height of that acquisition unit of that bearing position; The cell image at the sampling position of each acquisition unit is acquired using the acquisition focal height of each acquisition unit at each acquisition position.

2. The method according to claim 1, characterized in that, The imaging camera is located vertically below the objective lens; Determining the initial focal parameters for the 3D control platform to acquire data from the transparent carrier includes: The initial bearing position of the transparent carrier is moved above the objective lens by the horizontal motion platform. With both the bright field light source and the imaging camera turned on, the vertical axis motor is adjusted and multiple images of the transparent carrier surface at different focus heights are acquired. The focus height corresponding to the transparent carrier surface image with the highest clarity among the transparent carrier surface images is determined as the initial focal plane height. When the vertical axis motor is at the initial focal plane height, the laser source is turned on, the center position of the laser spot in the light spot image acquired by the imaging camera is recorded, and the initial focal parameters are obtained by combining the initial focal plane height and the center position of the laser spot.

3. The method according to claim 2, characterized in that, When traversing to the preset laser position of each acquisition unit at each carrier position, determining the acquisition focal height for bright-field photography at that carrier position at that acquisition unit includes: For each acquisition unit of each traversed bearing position, a laser image is acquired at the preset laser position with the laser source turned on, and the center position of the light spot in the laser image is determined. For each acquisition unit at each traversed carrier position, the center position of the laser spot at the preset laser position is calculated, and the deviation value between the center position of the laser spot and the latest laser spot center position is calculated. Based on the deviation value, the acquisition focal height of the acquisition unit at the carrier position for bright-field photography is calculated.

4. The method according to any one of claims 1-3, characterized in that, The carrier positions in the transparent carrier are arranged in a row and column matrix, the collection units in each carrier position are arranged in a row and column matrix, and all collection units in the transparent carrier are arranged in a row and column matrix. The method also includes: The horizontal motion platform is driven to move to the initial acquisition unit of the transparent carrier. When the horizontal motion platform moves to each acquisition unit, the cell imaging task of that acquisition unit is executed. After completing the cell imaging task of each acquisition unit, the horizontal motion platform is driven to move in the first horizontal direction until it reaches the next acquisition unit and performs its cell imaging task. After the horizontal motion platform moves to the last acquisition unit in the first horizontal direction and completes its cell imaging task, it is driven to move to the second horizontal direction to the adjacent acquisition unit that has not yet been imaged and to perform its cell imaging task. Then, the horizontal motion platform is driven to move in the opposite direction of the current first horizontal direction to continue to image the acquisition units in the current first horizontal direction one by one. After the horizontal motion platform moves to the last acquisition unit in the second horizontal direction, the cell imaging task of the transparent carrier ends when the cell imaging task of the last acquisition unit in the current first horizontal dimension is completed.

5. A cell sample imaging system, characterized in that, It includes a 3D control platform, a bright field light source, a laser light source, objective lenses, a transparent carrier, an imaging camera, and a controller; The three-dimensional control platform includes a horizontal motion platform and a vertical axis motor. The vertical axis motor is fixed with the objective lens. The transparent carrier is placed on the horizontal motion platform. The transparent carrier contains multiple support positions for culturing cells. The bright field light source emits a beam that passes through the transparent carrier and the objective lens in sequence and is then received by the imaging camera. The laser light source is located between the objective lens and the transparent carrier. The controller includes: A focal parameter initialization unit is used to determine the initial focal parameters for the three-dimensional control platform to collect data from the transparent carrier. The focal height determination unit is used to determine the focal height of the carrier for bright-field photography when, during the process of traversing each carrier position of the transparent carrier to perform bright-field photography using the initial focal parameters as the initial parameters for bright-field photography traversal, the carrier position is traversed to the preset laser position of each acquisition unit of each carrier position. A synchronous moving unit is used to drive the vertical axis motor to move to the acquisition focal height of the acquisition unit at the bearing position during the process of the horizontal motion platform moving from the preset laser position of each acquisition unit at each bearing position to the sampling position of the acquisition unit at that bearing position; An imaging unit is used to acquire a cell image of the carrier at the sampling position of the acquisition unit at the acquisition focal height of each acquisition unit of each carrier position.

6. The system according to claim 5, characterized in that, The imaging camera is located vertically below the objective lens; The focal parameter initialization unit includes: The first focal parameter initialization subunit is used to move the initial bearing position of the transparent carrier to above the objective lens via the horizontal motion platform. The second focal parameter initialization subunit is used to adjust the vertical axis motor and acquire multiple transparent carrier surface images with different focus heights when both the bright field light source and the imaging camera are turned on, and to determine the focus height corresponding to the transparent carrier surface image with the highest clarity among the transparent carrier surface images as the initial focal plane height. The third focal parameter initialization subunit is used to turn on the laser source when the vertical axis motor is at the initial focal plane height, record the center position of the laser spot in the light spot image acquired by the imaging camera, and combine the initial focal plane height and the center position of the laser spot to obtain the initial focal parameters.

7. The system according to claim 6, characterized in that, The focal height determination unit includes: The spot center determination unit is used to acquire a laser image at the preset laser position for each acquisition unit of each traversed bearing position under the condition that the laser source is turned on, and to determine the center position of the spot in the laser image. The focal height calculation unit is used to calculate the deviation between the center position of the laser spot and the latest center position of the laser spot for each acquisition unit at each traversed carrier position, and to calculate the focal height of the acquisition unit at the carrier position for bright-field photography based on the deviation value.

8. The system according to any one of claims 5-7, characterized in that, The carrier positions in the transparent carrier are arranged in a row and column matrix, the collection units in each carrier position are arranged in a row and column matrix, and all collection units in the transparent carrier are arranged in a row and column matrix. The controller also includes: The acquisition unit task execution unit is used to drive the horizontal motion platform to move to the initial acquisition unit of the transparent carrier. When the horizontal motion platform moves to each acquisition unit, it executes the cell imaging task of that acquisition unit. A one-dimensional horizontal movement unit is used to drive the horizontal motion platform to move in the first-dimensional horizontal direction after completing the cell imaging task of each acquisition unit, until it reaches the next acquisition unit of that acquisition unit and performs its cell imaging task. A two-dimensional horizontal movement unit is used to drive the horizontal motion platform to move in the second dimension to the adjacent acquisition unit that has not yet been imaged and to perform its cell imaging task after the horizontal motion platform moves in the first dimension horizontal direction to the last acquisition unit and completes its cell imaging task. It also drives the horizontal motion platform to move in the opposite direction of the current first dimension horizontal direction to continue to image the acquisition units in the current first dimension horizontal direction one by one. The imaging task completion unit is used to end the cell imaging task of the transparent carrier after the horizontal motion platform moves to the last acquisition unit in the second horizontal direction and completes the cell imaging task of the last acquisition unit in the current first horizontal dimension.

9. A cell sample imaging device, characterized in that, Including memory and processor; The memory is used to store programs; The processor is configured to execute the program to implement the various steps of the cell sample imaging method as described in any one of claims 1-4.

10. A storage medium having a computer program stored thereon, characterized in that, When the computer program is executed by the processor, it implements the various steps of the cell sample imaging method as described in any one of claims 1-4.

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