Method and device for optimizing living cells through cooperation of double visual fields

Abstract

The invention provides a dual-view collaborative living cell optimization method and equipment. The method comprises the following steps: calibrating a first view and a second view; performing activity evaluation on living cells in the culture dish under a first visual field, and screening first target living cells; adjusting the position of the culture dish, and moving the first target living cells into a second visual field; in a second visual field, performing form evaluation on the first target living cells, and screening out second target living cells; the position of the culture dish is adjusted, and the second target living cells are fixed through the laser spots. Through double-view calibration, the alignment precision of a laser spot and a living cell is improved, and the living cell is prevented from deviating from the laser spot, so that the living cell is prevented from struggling off due to infirm fixation, and the living cell is prevented from being damaged, low in grabbing efficiency and the like; living cells are moved into a second visual field by adjusting a culture dish, so that the form evaluation precision is improved; and through double screening of vitality and form, the misjudgment risk of a single evaluation dimension is reduced, and the reliability of a screening result is improved.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of live cell sorting technology, and particularly relates to a double-viewfield cooperative live cell optimization method and device. BACKGROUND

[0002] The viability and morphology of live cells (such as sperm and ova) are key parts of live cell examination, and are of great significance to reproductive and developmental research, especially to clinical reproductive medicine. Existing medical clinical detection uses different sample preparation and microscopic detection methods to respectively detect the concentration, viability and morphology of live cells.

[0003] However, in the existing morphology detection and analysis process, since live cells are usually in motion, and the motion speed of some live cells is extremely fast, clear imaging cannot be achieved, resulting in low accuracy of morphology detection results. SUMMARY

[0004] The present application provides a double-viewfield cooperative live cell optimization method and device to solve the technical problem of low accuracy of morphology detection caused by the motion of live cells in the prior art.

[0005] According to a first aspect of the present application, a double-viewfield cooperative live cell optimization method is provided, which comprises: calibrating the field center of a first viewfield and the field center of a second viewfield, so that the field center of the first viewfield coincides with the field center of the second viewfield; under the first viewfield, performing viability evaluation on each live cell in a target region corresponding to the first viewfield in a culture dish to screen at least one first target live cell; for each first target live cell in the target region, adjusting the position of the culture dish to move the first target live cell into the field range of the second viewfield; under the second viewfield, performing morphology evaluation on each first target live cell to screen at least one second target live cell; according to the position of each second target live cell in the target region, adjusting the position of the culture dish to make the laser spot emitted by a laser fixed on the corresponding second target live cell in turn and transfer the second target live cell, the position of the laser spot coinciding with the center of the cell imaging region of the second viewfield; adjusting the position of the culture dish to make different regions of the culture dish correspond to the first viewfield respectively, and repeating the viability evaluation on each live cell in the target region until the fixation and transfer of all second target live cells in the culture dish are completed.

[0006] In some embodiments, the calibrating the field center of the first field of view and the field center of the second field of view comprises: selecting one of the living cells in the culture dish; fixing the selected living cell by a laser spot emitted by the laser under the second field of view; and adjusting the field center of the first field of view based on the position of the laser spot so that the field center of the first field of view coincides with the position of the laser spot.

[0007] In some embodiments, the adjusting the field center of the first field of view based on the position of the laser spot comprises: magnifying the field center area of the first field of view to determine the field center of the first field of view; and adjusting the field center of the first field of view based on the position of the laser spot so that the field center of the first field of view coincides with the position of the laser spot.

[0008] In some embodiments, before fixing the selected living cell by the laser spot emitted by the laser, the method further comprises: adjusting the position of the laser so that the position of the laser spot emitted by the laser coincides with the field center of the second field of view.

[0009] In some embodiments, the evaluating the viability of each living cell in the target region corresponding to the first field of view in the culture dish under the first field of view to screen at least one first target living cell comprises: collecting a plurality of motion images of a plurality of living cells in the target region; determining a motion parameter of each living cell in the target region according to the plurality of motion images of the plurality of living cells in the target region; and selecting a living cell with a speed greater than a first threshold value as a first target living cell according to the motion parameter of each living cell in the target region.

[0010] In some embodiments, the evaluating the morphology of each first target living cell in the target region under the second field of view to screen at least one second target living cell comprises: collecting a plurality of morphology images of each first target living cell in the target region; determining a morphology parameter of each first target living cell according to the plurality of morphology images of each first target living cell; and selecting a first target living cell with a morphology parameter meeting a second threshold value as a second target living cell according to the morphology parameter of each first target living cell.

[0011] In some embodiments, the field of view range of the first field of view is greater than the field of view range of the second field of view.

[0012] In some embodiments, the method includes: grasping the second target living cell and transferring it to a target location while the second target living cell is fixed in place.

[0013] According to a second aspect of this application, a dual-field-of-view collaborative live cell selection device is provided, characterized in that it comprises: a first microscopic imaging system having a first field of view and used to assess the viability of each live cell in a culture dish under the first field of view to screen for at least one first target live cell; a position adjustment mechanism for supporting the culture dish and being able to adjust the position of the culture dish; a second microscopic imaging system having a second field of view and used to assess the morphology of each first target live cell under the second field of view to screen for at least one second target live cell; and an optical tweezers module including a laser for emitting a laser spot to fix the live cells in the culture dish.

[0014] In some embodiments, the optical tweezers module further includes a laser adjustment unit for adjusting the intensity of the laser spot.

[0015] In summary, the dual-field collaborative live cell optimization method and apparatus provided in this application have at least the following beneficial effects: In this application, before assessing the viability and morphology of live cells in the culture dish, the centers of the first and second fields of view are calibrated. This ensures that the centers of the first and second fields of view coincide, improving the alignment accuracy between the laser spot and the second target live cells when the laser beam is used to fix them. This prevents the second target live cells from deviating from the laser spot, which could lead to poor fixation and the cells escaping, resulting in low subsequent grasping efficiency. After calibrating the centers of the first and second fields of view, viability assessment is performed on each live cell in the culture dish under the first field of view to select at least one first target live cell, and morphological assessment is performed on each first target live cell under the second field of view to select at least one second target live cell. This achieves quality screening of live cells. Thus, by using both viability and morphology as dual criteria for screening, the risk of misjudgment based on a single assessment dimension is reduced, improving the reliability of the final screening results.

[0016] Furthermore, in order to improve the accuracy of morphological assessment of each first target live cell under the second field of view, the position of the culture dish is adjusted before morphological assessment of each first target live cell to move the first target live cell into the field of view of the second field of view. This greatly improves the accuracy of morphological assessment when assessing the first target live cell.

[0017] Furthermore, in the entire process of dual-field collaborative live cell selection, the positions and ranges of the first and second fields of view remain unchanged. Instead, the position of the culture dish is adjusted to match the center of the first and second fields of view, thereby ensuring that each live cell can be evaluated for viability and morphology under the same conditions. This avoids errors introduced by changes in the field of view, thereby improving the consistency of evaluation of all live cells and helping to reduce the risk of screening errors. Attached Figure Description

[0018] To more clearly illustrate the technical solutions in the specific embodiments of this application, the drawings used in the specific embodiments will be briefly introduced below. Obviously, the drawings described below are some embodiments of this application. For those skilled in the art, other drawings can be obtained from these drawings without creative effort.

[0019] Figure 1 This is a flowchart illustrating a dual-field collaborative live cell selection method provided in one embodiment of this application. Figure 2 A flowchart illustrating a dual-field collaborative live cell selection method provided in another embodiment of this application; Figure 3 A flowchart illustrating a dual-field collaborative live cell selection method provided in another embodiment of this application; Figure 4 This is a schematic diagram of a dual-field collaborative live cell selection device provided for an embodiment of this application. Detailed Implementation

[0020] To make the above and other features and advantages of this application clearer, the application is further described below with reference to the accompanying drawings. It should be understood that the specific embodiments given herein are for the purpose of explanation to those skilled in the art, and are exemplary only, not restrictive.

[0021] In the following description, numerous specific details are set forth to provide a thorough understanding of this application. However, it will be apparent to those skilled in the art that the specific details are not required to practice this application. In other instances, well-known steps or operations have not been described in detail to avoid obscuring this application.

[0022] The dual-field collaborative live cell selection method provided in this application embodiment can be implemented by the dual-field collaborative live cell selection device provided in this application embodiment.

[0023] refer to Figure 1 This application provides a dual-field collaborative method for selecting live cells, the method comprising: Step 101: Calibrate the visual field center of the first visual field and the visual field center of the second visual field so that the visual field center of the first visual field coincides with the visual field center of the second visual field.

[0024] Understandably, the purpose of calibrating the visual field centers of the first and second visual fields is to make them coincide. Calibration can be performed manually or automatically through system settings.

[0025] The dual-field collaborative live cell selection method provided in this application involves two fields of view: a first field of view and a second field of view. In the dual-field collaborative live cell selection process, image data acquired under the first and second fields of view can be used to screen the quality of somatic cells. The first and second fields of view have different field of view ranges, which can be achieved by selecting microscopic imaging systems with different field of view ranges. Furthermore, the field of view range of the first field of view is larger than that of the second field of view.

[0026] Understandably, the field of view of an image is inversely proportional to the magnification of the microscopic imaging system. Low magnification can achieve a large field of view, but not high resolution. High magnification can achieve high resolution, but cannot directly achieve a large field of view. In other words, the first field of view is a large field of view with low resolution, and the second field of view is a small field of view with high resolution. Obviously, a large field of view with low resolution is more conducive to the analysis of the motion of living cells, while a small field of view with high resolution is more conducive to the analysis of the morphology of living cells.

[0027] Step 102: Under the first field of view, assess the viability of each live cell in the target area corresponding to the first field of view in the culture dish to screen out at least one first target live cell.

[0028] A sufficient number of live cells can be pre-cultured or placed in the culture dish. The culture dish and the live cells inside can be placed on the stage before calibrating the center of the first field of view and the center of the second field of view. In this way, the live cells in the culture dish can be used for positioning to calibrate the center of the first field of view and the center of the second field of view. This will be explained in detail below.

[0029] Understandably, the first field of view can be a low-power or wide-angle field of view, while the second field of view can be a high-power or a field of view with a specific function. To ensure image acquisition accuracy, the first field of view is usually smaller than the area of ​​the culture dish. Therefore, the viability of live cells in different regions within the culture dish can be assessed separately under the first field of view, thereby achieving the viability assessment of all live cells in the culture dish. Furthermore, by moving the position of the culture dish, different regions of the culture dish can be aligned with (i.e., covered by) the first field of view. When a certain region of the culture dish aligns with the first field of view, the viability of live cells within that region can be assessed.

[0030] Specifically, when assessing the viability of each living cell in a culture dish, image processing and target location tracking techniques can be used for evaluation, and this application does not impose any specific limitations.

[0031] Step 103: For each first target living cell within the target area, adjust the position of the culture dish to move the first target living cell into the field of view of the second field of view.

[0032] It should be noted that, since living cells are usually in motion, if a first target living cell is not within the field of view of the second field of view before morphological evaluation, morphological evaluation cannot be performed or will be affected. By adjusting the position of the culture dish, the first target living cell can be moved as a whole into the field of view of the second field of view, thus facilitating morphological evaluation. Preferably, by adjusting the position of the culture dish, the first target living cell is placed at the center of the cell imaging area of ​​the second field of view, which helps to improve the accuracy of morphological evaluation of the first living cell. Here, the cell imaging area of ​​the second field of view refers to a region of a given size defined with any position within the second field of view as the center point, and cell images are acquired through this given region; this given region is called the cell imaging area.

[0033] To better understand this solution, an example is provided to illustrate how to adjust the position of the petri dish.

[0034] Obtain the center of the cell imaging region in the second field of view and determine the coordinates as the origin (0,0); Real-time acquisition of the relative position of the first target living cell with respect to the origin and determination of the coordinates as (X,Y); The culture dish is moved in the opposite direction by X and Y distances until the coordinates of the first target living cell approach (0,0). The direction from the origin to the first target living cell is considered positive; therefore, the reverse movement is in the opposite direction to the positive direction.

[0035] It should be noted that living cells are in motion and their positions are constantly changing. Adjusting the position of the culture dish is mainly to track the living cells and ensure that the primary target living cell is centered in the cell imaging area of ​​the second field of view.

[0036] It should be understood that the size and resolution (pixels) of the cell imaging region in the second field of view are known. The center position of the cell imaging region in the second field of view can be easily determined based on the size of the image and the number of pixels. A two-dimensional coordinate system is constructed with the center position of the cell imaging region as the origin, that is, mutually perpendicular X-axis and Y-axis are constructed in the imaging plane. The coordinates of the first target living cell can be determined based on the pixel difference between it and the origin, that is, the pixel difference between the X-axis and the pixel difference between the Y-axis.

[0037] The movement of the petri dish can be controlled by either a synchronized movement or a single-step movement. The synchronized movement means that the X and Y coordinates are adjusted synchronously, while the single-step movement means that either the X-axis position or the Y-axis position is adjusted first, and then the other position is adjusted. The synchronized movement method is preferred.

[0038] Step 104: Under the second field of view, perform morphological evaluation on each first target live cell to screen out at least one second target live cell.

[0039] Specifically, morphological evaluation of each first target living cell can be performed using image processing and analysis techniques, which are not specifically limited in this application.

[0040] In this process, by performing morphological evaluation on each first target live cell, the head shape and size, tail shape and size, vacuole position and size, etc. of each first target live cell can be determined. Then, by comparing the head shape and / or size, tail shape and / or size, vacuole position and / or size of the first target live cell with the corresponding target image, the second target live cell can be screened out.

[0041] Step 105: Adjust the position of the culture dish according to the position of each second target living cell in the target area so that the laser spot emitted by the laser sequentially fixes the corresponding second target living cells and transfers the second target living cells, and the position of the laser spot coincides with the center of the cell imaging area of ​​the second field of view.

[0042] The position of each second target live cell can be acquired in real time under the first field of view. After the second target live cells are selected, the position of the culture dish can be adjusted according to the position of each second target live cell to move the second target live cell to the center of the cell imaging area of ​​the second field of view. At this time, the laser is turned on, and the laser spot emitted by the laser can be directed towards the second target live cell to fix the second target live cell.

[0043] Step 106: Adjust the position of the culture dish so that different areas of the culture dish correspond to the first field of view, and repeat the viability assessment of each live cell in the target area until the fixation and transfer of all second target live cells in the culture dish are completed.

[0044] Specifically, repeat steps 102 to 105 above to complete the viability assessment analysis of live cells in each region of the culture dish, and sequentially identify second target live cells that meet the conditions in each region of the culture dish, and complete the fixation and transfer of these second target live cells. It should be understood that the second target live cells here refer to live cells that meet both viability and morphological requirements.

[0045] Understandably, by traversing all the live cells in the culture dish, the motility parameters of all live cells within the dish can be determined. During the traversal, operations such as numbering the live cells can be performed to avoid duplicate determinations that would affect screening efficiency.

[0046] Therefore, in this embodiment, before assessing the viability and morphology of live cells in the culture dish, the centers of the first and second fields of view are calibrated. This ensures that the centers of the first and second fields of view coincide, improving the alignment accuracy between the laser spot and the second target live cells when the laser spot is used to fix them. This prevents the second target live cells from deviating from the laser spot, which could lead to poor fixation and the cells escaping the laser spot, resulting in low subsequent grasping efficiency. After calibrating the centers of the first and second fields of view, viability assessment is performed on each live cell in the culture dish under the first field of view to select at least one first target live cell, and morphology assessment is performed on each first target live cell under the second field of view to select at least one second target live cell. This achieves quality screening of live cells. Thus, by using both viability and morphology as dual criteria for screening, the risk of misjudgment based on a single assessment dimension is reduced, improving the reliability of the final screening results.

[0047] Furthermore, in order to improve the accuracy of morphological assessment of each first target live cell under the second field of view, the position of the culture dish is adjusted before morphological assessment of each first target live cell to move the first target live cell into the field of view of the second field of view. This greatly improves the accuracy of morphological assessment when assessing the first target live cell.

[0048] Furthermore, in the entire process of dual-field collaborative live cell selection, the positions and ranges of the first and second fields of view remain unchanged. Instead, the position of the culture dish is adjusted to adapt to the first and second fields of view, thereby ensuring that each live cell can be evaluated for viability and morphology under the same conditions. This avoids errors introduced by changes in the field of view, thereby improving the consistency of evaluation of all live cells and helping to reduce the risk of screening errors.

[0049] refer to Figure 2 The dual-field collaborative live cell optimization method provided in this application embodiment may include the following steps: Step 201: Select one of the living cells in the culture dish.

[0050] Step 202: Adjust the position of the laser so that the position of the laser spot emitted by the laser coincides with the center of the field of view of the second field of view.

[0051] Step 203: Under the second field of view, the selected living cells are fixed by a laser spot emitted by the laser.

[0052] Step 204: Based on the position of the laser spot, adjust the center of the first field of view so that the center of the first field of view coincides with the position of the laser spot.

[0053] In other words, this application first selects one of the living cells in the culture dish and uses it as a reference. Then, under the second field of view, the position of the laser is adjusted to change the position of the laser spot under the second field of view, so that the laser spot coincides with the center of the field of view of the second field of view. The selected living cell is fixed by the laser spot emitted by the laser. Then, based on the position of the laser spot, the center of the field of view of the first field of view is adjusted so that the center of the field of view of the first field of view coincides with the position of the laser spot. That is, the center of the field of view of the first field of view, the center of the field of view of the second field of view, and the position of the laser spot coincide, thereby achieving calibration and adjustment of the center of the field of view of the first and second fields of view.

[0054] Thus, through steps 201-204, by utilizing the position of any living cell in the culture dish and adjusting the laser spot position, the mechanical displacement or optical parameters of the first field of view are adjusted so that the center of the first field of view coincides with the laser spot, completing the dual-field-of-view center calibration. This adjustment method eliminates the need for manual focusing, reduces reliance on operator experience, and significantly improves the positioning consistency in multi-field operations.

[0055] Furthermore, before fixing the selected living cells with the laser spot emitted by the laser, the position of the laser can be adjusted so that the position of the laser spot emitted by the laser coincides with the center of the field of view of the second field of view.

[0056] Furthermore, in order to improve positioning accuracy, the central region of the first field of view can be magnified first to determine the center of the first field of view; then, based on the position of the laser spot, the center of the first field of view is adjusted so that the center of the first field of view coincides with the position of the laser spot.

[0057] By magnifying the central region of the first field of view, the center of the first field of view can be accurately determined, reducing the positioning error of the field of view center. This helps to improve the positioning accuracy of the center of the first field of view and the laser spot.

[0058] Step 205: Under the first field of view, assess the viability of each live cell in the target area corresponding to the first field of view in the culture dish to screen out at least one first target live cell.

[0059] Step 206: For each first target live cell within the target area, adjust the position of the culture dish to move the first target live cell into the field of view of the second field of view. Understandably, adjusting the position of the culture dish is equivalent to adjusting the position of the first target live cell.

[0060] Step 207: In the second field of view, perform morphological evaluation on each first target live cell to screen out at least one second target live cell.

[0061] Step 208: Adjust the position of the culture dish according to the position of each second target living cell in the target area so that the laser spot emitted by the laser sequentially fixes the corresponding second target living cells and transfers the second target living cells, and the position of the laser spot coincides with the center of the cell imaging area of ​​the second field of view.

[0062] In addition, while the second target living cell is fixed, the second target living cell can also be grasped and transferred to the target location, which is not specifically limited in this application.

[0063] In this embodiment, a single live cell is first selected from the culture dish and fixed using a laser spot. Then, by adjusting the position or optical parameters of the first field of view, the center of the first field of view can be aligned with the laser spot. This process eliminates the need for manual focusing, significantly reducing reliance on operator experience. Next, under the first field of view, the viability of each live cell in the culture dish is assessed to screen for at least one first target live cell. This allows for the screening of live cells with varying viability, identifying those within the required viability range as the first target live cells. Subsequently, the first target live cells are moved to the second field of view, where microscopic morphological analysis identifies first target live cells with morphologically suitable ones as second target live cells. Thus, rapid initial screening using the first field of view followed by precise rescreening using the second field of view, based on phased evaluation and combining macroscopic and microscopic characteristics, improves the comprehensiveness of the screening, making it suitable for high-throughput cell screening scenarios. Furthermore, between the initial and rescreening stages, the position of each first target cell is individually adjusted to ensure it remains within the second field of view, thereby avoiding evaluation bias caused by field of view shifts.

[0064] refer to Figure 3 The dual-field collaborative live cell optimization method provided in this application embodiment may include the following steps: Step 301: The visual field center of the first visual field and the visual field center of the second visual field are calibrated so that the visual field center of the first visual field coincides with the visual field center of the second visual field.

[0065] Step 302: Under the first field of view, acquire multiple consecutive motion images of multiple living cells in the target area corresponding to the first field of view of the culture dish.

[0066] Understandably, since living cells are usually in motion, the multiple consecutive motion images of multiple living cells within the target area of ​​the culture dish acquired in this application refer to the continuous capture of multiple motion images (i.e., frame-by-frame images) within a certain time period, with each motion image containing all living cells within the target area. These multiple consecutive motion images can be used to determine the positions of multiple living cells at different times, thereby determining the movement speed of each living cell based on its position at different times.

[0067] Step 303: Determine the motion parameters of each living cell in the target area based on multiple motion images of multiple living cells in the target area.

[0068] The motion parameters of each living cell include position and velocity. After acquiring multiple motion images of multiple living cells within the target area, image recognition technology can be used to identify the position of each living cell at the corresponding moment in each motion image, so as to obtain the continuous motion trajectory of the living cell during the image acquisition time. Then, based on the motion trajectory and motion time of the living cell (i.e., the time of acquiring multiple motion images), the motion parameters (such as motion velocity) of each living cell can be calculated.

[0069] Step 304: Based on the motion parameters of each living cell in the target area, select all living cells with a velocity greater than the first threshold as the first target living cells.

[0070] Understandably, the first target living cell can be some or all of the multiple living cells in the target area. It can be selected according to specific screening conditions. For example, living cells whose average velocity or highest instantaneous velocity of each living cell within a certain range during the image acquisition time can be selected as the first target living cell. This application does not limit this.

[0071] Step 305: For each first target live cell, adjust the position of the culture dish to move the first target live cell into the field of view of the second field of view.

[0072] Specifically, the position adjustment of the culture dish can be accomplished through robotic arms, multi-axis motion mechanisms, or automated equipment.

[0073] Step 306: Under the second field of view, acquire multiple sets of morphological images of each first target living cell within the target area.

[0074] Each morphological image contains, but is not limited to, at least one of the head, tail, and vacuoles of the first target living cell.

[0075] Step 307: Determine the morphological parameters of each first target living cell based on the multiple sets of morphological images of each first target living cell collected.

[0076] Step 308: Based on the morphological parameters of each first target living cell, select the first target living cells whose morphological parameters meet the second threshold as the second target living cells.

[0077] Step 309: Adjust the position of the culture dish according to the position of the second target living cells so that the laser spot emitted by the laser fixes the second target living cells. The position of the laser spot coincides with the center of the second field of view.

[0078] Step 310: While the second target living cell is fixed, grasp the second target living cell and transfer it to the target location.

[0079] It should be noted that the above examples are merely illustrative and should not be construed as limiting the determination of second target living cells, etc., in the embodiments of this application.

[0080] Therefore, in this embodiment, the combination of steps 301, 309, and 310 can improve the alignment accuracy between the laser spot and the second target living cell, preventing the second target living cell from deviating from the laser spot and causing it to break free due to insufficient fixation, thus leading to problems such as cell damage, low grasping efficiency, and low grasping accuracy. Through dynamic image acquisition and motion parameter analysis (such as velocity threshold screening) in steps 302-304, highly active cells can be accurately identified. Furthermore, capturing cell movement trajectories in multiple consecutive frames and combining this with velocity calculations (such as average velocity or instantaneous velocity) can effectively exclude low-activity or stationary cells, improving the accuracy of the initial screening. Through secondary screening using morphological parameters (such as head, tail, and vacuolar features) in steps 306-308, a composite evaluation method combining motion and morphological parameters is formed, improving the comprehensiveness of the screening and identifying high-quality living cells. For example, in this embodiment, high-speed moving cells may be rejected due to morphological abnormalities (such as excessive vacuolarity), avoiding misjudgments caused by a single indicator.

[0081] refer to Figure 4 This application provides a dual-field collaborative live cell optimization device, which includes a first microscopic imaging system 410, a position adjustment mechanism 420, a second microscopic imaging system 430, and an optical tweezers module 440.

[0082] The first microscopic imaging system 410 has a first field of view and is used to assess the viability of each living cell in the culture dish under the first field of view in order to screen out at least one first target living cell.

[0083] Specifically, the first microscopic imaging system 410 may include a first image acquisition module, a first analysis module, and a first determination module. The first image acquisition module is used to acquire multiple consecutive motion images of multiple living cells within a target area corresponding to the first field of view in a culture dish. The first analysis module is used to determine the motion parameters of each living cell within the target area based on the multiple motion images of the living cells within the target area. The first determination module is used to select all living cells with a velocity greater than a first threshold as the first target living cells based on the motion parameters of each living cell in the culture dish.

[0084] The position adjustment mechanism 420 is used to support the culture dish and adjust its position. For example, during the viability assessment of live cells in the culture dish, the position adjustment mechanism 420 can be used to adjust the position of the culture dish so that different areas of the culture dish correspond to the first field of view, and enable the first image acquisition module, the first analysis module, and the first determination module to sequentially repeat multiple motion images of multiple live cells in the target area corresponding to the first field of view of the culture dish until the motion parameters of all live cells in the culture dish are determined; before performing morphological assessment on each first target live cell, the position adjustment mechanism 420 can be used to adjust the position of the culture dish to move the first target live cell into the field of view of the second field of view; after performing morphological assessment on each first target live cell, the position adjustment mechanism 420 can adjust the position of the culture dish according to the position of each second target live cell so that the laser spot emitted by the laser fixes the second target live cell.

[0085] Specifically, a stage can be provided on the position adjustment mechanism 420, and the petri dish is placed on the stage. The position adjustment mechanism 420 can be a robotic arm, a multi-axis motion mechanism, or an automated device, etc., and this application does not make any specific limitation.

[0086] The second microscopic imaging system 430 has a second field of view and is used to perform morphological evaluation on each first target live cell under the second field of view in order to screen out at least one second target live cell.

[0087] Specifically, the second microscopic imaging system 430 may include a second image acquisition module, a second analysis module, and a second determination module. The second image acquisition module is used to acquire multiple sets of morphological images of each first target living cell under a second field of view; the second analysis module is used to determine the morphological parameters of each first target living cell based on the acquired multiple sets of morphological images; and the second determination module is used to select all first target living cells whose morphological parameters meet a second threshold as second target living cells based on the morphological parameters of each first target living cell.

[0088] The optical tweezers module 440 includes a laser 441, which emits a laser spot to immobilize living cells in a culture dish.

[0089] In this embodiment, based on the first microscopic imaging system 410, the position adjustment mechanism 420, the second microscopic imaging system 430, and the optical tweezers module 440, before evaluating each live cell in the culture dish, the position adjustment mechanism 420 and the optical tweezers module 440 can use the laser 441 of the optical tweezers module 440 to emit a laser spot to fix any selected live cell, and in the second field of view, the laser spot is aligned with the center of the field of view of the second field of view, and the live cell is fixed. Then, based on the position of the laser spot, the center of the field of view of the first field of view is adjusted so that the center of the field of view of the first field of view is aligned with the position of the laser spot. This allows for the calibration of the center of the field of view of the first field of view and the center of the field of view of the second field of view, so that the center of the field of view of the first field of view is aligned with the center of the field of view of the second field of view. Then, the viability of each live cell in the culture dish is assessed by the first microscopic imaging system 410 to screen out the first target live cells. Next, for each first target live cell, the position of the culture dish is adjusted by the position adjustment mechanism 420 to move the first target live cell into the field of view of the second field of view. After that, the morphology of each first target live cell is assessed by the second microscopic imaging system 430 to screen out the second target live cells. Then, according to the position of each second target live cell, the position of the culture dish is adjusted again by the position adjustment mechanism 420 so that the laser spot emitted by the laser 441 can fix the second target live cells, which facilitates the subsequent transfer of the second target live cells, thereby completing the high-quality screening of live cells in the culture dish.

[0090] Therefore, in the dual-field-of-view collaborative live cell selection device of this application embodiment, before evaluation, the center of the first field of view and the center of the second field of view are calibrated through the position adjustment mechanism 420 and the optical tweezers module 440. This improves the alignment accuracy between the laser spot and the second target live cell, preventing the second target live cell from deviating from the position of the laser spot, which could lead to the live cell escaping the laser spot due to insecure fixation, resulting in problems such as live cell damage, low grasping efficiency, and low grasping accuracy. Furthermore, the first microscopic imaging system 410 performs rapid initial screening, and the second microscopic imaging system 430 performs precise secondary screening. Based on staged evaluation and combining macroscopic and microscopic features, the comprehensiveness of the screening is improved, making it suitable for high-throughput cell screening scenarios. In addition, between the initial screening and secondary screening, the position of each first target cell is individually adjusted by the position adjustment mechanism 420 to ensure it is within the second field of view, thereby avoiding evaluation deviations caused by field of view offset. In some embodiments, the optical tweezers module 440 includes a laser 441 and a laser adjustment unit 442. The laser 441 is used to emit a laser spot to fix live cells in a culture dish, and the laser adjustment unit 442 is used to adjust the intensity of the laser spot.

[0091] The laser adjustment unit 442 can adjust the intensity of the laser beam emitted by the laser, thereby adjusting the intensity of the laser spot on the living cells. This can avoid problems such as the laser spot being too strong and damaging the living cells, or the laser spot being too weak and not being able to fix the living cells firmly.

[0092] It should be understood that the specific features, operations, and details described above with respect to the method of this application can also be similarly applied to the device of this application, or vice versa. Furthermore, each step of the method of this application described above can be implemented by a corresponding component or unit of the device of this application.

[0093] It should be understood that the various modules / units of the device of this application can be implemented wholly or partially through software, hardware, firmware, or a combination thereof. Each module / unit can be embedded in the processor of the electronic device in hardware or firmware form or independent of the processor, or it can be stored in the memory of the electronic device in software form for the processor to call to execute the operation of each module / unit. Each module / unit can be implemented as an independent component or module, or two or more modules / units can be implemented as a single component or module.

[0094] The technical features described above can be combined arbitrarily. Although not all possible combinations of these technical features are described, any combination of these technical features should be considered to be covered by this specification, provided that such combination does not contain contradictions.

[0095] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of this application, and are not intended to limit them. Although this application has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some or all of the technical features therein. Such modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of this application.

Claims

1. A dual-field collaborative method for selecting optimal live cells, characterized in that, include: The visual field centers of the first and second visual fields are calibrated to make the visual field centers of the first and second visual fields coincide. Under the first field of view, the viability of each live cell in the target area corresponding to the first field of view in the culture dish is evaluated to screen out at least one first target live cell; For each of the first target living cells within the target area, the position of the culture dish is adjusted to move the first target living cell into the field of view of the second field of view; In the second field of view, each of the first target live cells is morphologically evaluated to screen out at least one second target live cell; Based on the position of each second target living cell within the target area, the position of the culture dish is adjusted so that the laser spot emitted by the laser sequentially fixes the corresponding second target living cells and transfers the second target living cells, and the position of the laser spot coincides with the center of the cell imaging area of ​​the second field of view; Adjust the position of the culture dish so that different areas of the culture dish correspond to the first field of view, and repeat the viability assessment of each live cell in the target area until the fixation and transfer of all the second target live cells in the culture dish are completed.

2. The dual-field synergistic live cell selection method according to claim 1, characterized in that, The calibration of the visual field center of the first visual field and the visual field center of the second visual field includes: Select one of the living cells from the culture dish; In the second field of view, the selected living cells are fixed by a laser spot emitted by a laser. Based on the position of the laser spot, the center of the first field of view is adjusted so that the center of the first field of view coincides with the position of the laser spot.

3. The dual-field synergistic live cell selection method according to claim 2, characterized in that, The adjustment of the center of the first field of view based on the position of the laser spot includes: The central region of the first field of view is magnified to determine the center of the first field of view; Based on the position of the laser spot, the center of the first field of view is adjusted so that the center of the first field of view coincides with the position of the laser spot.

4. The dual-field synergistic live cell selection method according to claim 2, characterized in that, Before fixing the selected living cells with a laser spot emitted by a laser, the method further includes: Adjust the position of the laser so that the position of the laser spot emitted by the laser coincides with the center of the field of view of the second field of view.

5. The dual-field synergistic live cell selection method according to claim 1, characterized in that, The step of assessing the viability of each live cell within the target region corresponding to the first field of view in the culture dish, under the first field of view, to screen out at least one first target live cell, includes: Acquire multiple consecutive motion images of multiple living cells within the target area; Based on the multiple motion images of the multiple living cells in the target area, determine the motion parameters of each living cell in the target area; Based on the motion parameters of each living cell within the target area, the living cells with a velocity greater than a first threshold are selected as the first target living cells.

6. The dual-field synergistic live cell selection method according to claim 1, characterized in that, The step of performing morphological evaluation on each of the first target living cells within the target region under the second field of view to screen out at least one second target living cell includes: Collect multiple sets of morphological images of each of the first target living cells within the target area; Based on the multiple sets of morphological images of each of the first target living cells, determine the morphological parameters of each of the first target living cells; Based on the morphological parameters of each first target living cell, the first target living cells whose morphological parameters meet the second threshold are selected as the second target living cells.

7. The dual-field synergistic live cell selection method according to claim 1, characterized in that, The field of view of the first field of view is larger than that of the field of view of the second field of view.

8. The dual-field synergistic live cell selection method according to any one of claims 1-7, characterized in that, The method includes: While the second target living cell is fixed in place, the second target living cell is grasped and transferred to the target location.

9. A dual-field collaborative live cell selection device, characterized in that, include: A first microscopic imaging system having a first field of view and used to assess the viability of each living cell in a culture dish under the first field of view in order to screen out at least one first target living cell; A position adjustment mechanism is used to support the culture dish and can adjust the position of the culture dish; A second microscopic imaging system has a second field of view and is used to perform morphological assessment on each of the first target live cells in the second field of view to screen out at least one second target live cell. as well as The optical tweezers module includes a laser for emitting laser beams to immobilize living cells in a culture dish.

10. The dual-field collaborative live cell optimization device according to claim 9, characterized in that, The optical tweezers module also includes a laser adjustment unit, which is used to adjust the intensity of the laser spot.

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