Imaging method and device, electronic equipment and storage medium

Abstract

The invention discloses an imaging method and device, electronic equipment and a storage medium. The imaging method comprises the following steps: an aperture diaphragm performs preliminary image acquisition of a sample on at least one hole site in a hole plate device by matching a preset aperture value with an imaging module; calculating a brightness difference value between the central position in the hole site and other different positions according to a preliminary image acquired by the preliminary image acquisition; the actual aperture value of the aperture diaphragm is correspondingly adjusted according to the brightness difference value so as to cooperate with the imaging module to perform actual image acquisition of the sample at each position in the hole site. Full-hole photographing is firstly completed through the aperture diaphragm according to the preset aperture value, the brightness difference between the center position and other different positions is obtained, and the actual aperture value of the aperture diaphragm is adjusted according to the brightness difference during next photographing, so that the other different positions can have large-angle light rays to form a counteracting effect with the concave liquid surface shape; the problem of edge blackening caused by a concave liquid surface formed by liquid surface tension is solved, and loss of cells in an edge area during imaging is reduced.

Description

Technical Field

[0001] This application relates to the field of optical imaging technology, and in particular to an imaging method, apparatus, electronic device and storage medium. Background Technology

[0002] Well plates, as sample carriers, are used in microscopic imaging scenarios. The samples inside the well plates are observed and analyzed through a microscopic imaging system to obtain relevant data about the samples.

[0003] However, during the actual imaging process of the well plate, due to the surface tension of the liquid, the sample liquid inside the well plate will form a concave liquid surface. Affected by the light absorption characteristics of the liquid, the brightness of the edge of the concave liquid surface will be lower than that of the central area, which can easily lead to the loss of cells in the edge area in the imaging image, affecting the quality of whole-well imaging and the accuracy of subsequent analysis.

[0004] To address the imaging defects caused by concave liquid surfaces, some related technologies have proposed designing apertures of different shapes to reshape the light and compensate for the influence of concave liquid surfaces. However, this method can only compensate for concave liquid surfaces of specific shapes, and since the size of the concave liquid surface is directly related to the amount of sample added to the orifice plate, this method only works well under specific conditions and has poor versatility.

[0005] Some related technologies compensate for concave liquid surfaces by manipulating the consumable material and using its shape. However, this method requires the development of new consumable materials, and using uncommon and mainstream consumable materials results in high costs.

[0006] Some related technologies address the concave liquid surface problem by adding a variable aperture between the imaging objective and the tube end, but this method actually reduces the numerical aperture of the objective, thereby lowering the resolution of the imaging system. Summary of the Invention

[0007] This application proposes an imaging method to effectively solve the technical problems such as imaging defects caused by concave liquid surfaces during imaging of orifice plates in related technologies.

[0008] The first aspect of this application provides an imaging method, including the following steps:

[0009] The aperture stop, with a preset aperture value, works in conjunction with the imaging module to acquire a preliminary image of the sample at at least one well in the orifice plate device.

[0010] Calculate the brightness difference between the center position and other different positions within the aperture based on the preliminary image obtained from the preliminary image acquisition.

[0011] The actual aperture value of the aperture stop is adjusted according to the brightness difference to cooperate with the imaging module to acquire actual images of the sample at each position within the aperture.

[0012] Furthermore, the preliminary image acquisition of the sample includes:

[0013] Select the target aperture to be imaged, take multiple images of the target aperture, and fuse the images after the imaging area covers the cross-sectional area of ​​the aperture to obtain the preliminary image.

[0014] Further, the step of calculating the brightness difference between the center position within the aperture and other different positions based on the preliminary image obtained from the preliminary image acquisition includes:

[0015] Based on the image brightness value, the preliminary image obtained by fusion is divided into regions to obtain a central region and an edge region. The difference between the image brightness value of the central region and the image brightness value of the edge region is calculated to obtain the brightness difference value.

[0016] Furthermore, the step of acquiring actual images of the sample at each location within the aperture, in conjunction with the imaging module, includes:

[0017] Actual image acquisition is performed based on the central region and the edge region respectively;

[0018] When the central region is actually image acquired, the aperture stop cooperates with the imaging module to acquire the image using the preset aperture value.

[0019] When the edge region is actually captured, the aperture stop is adjusted to the target brightness value based on the brightness value of the central region during imaging. After the actual brightness value of the edge region reaches the target brightness value, the image is captured in conjunction with the imaging module.

[0020] Further, adjusting the actual aperture value of the aperture stop according to the brightness difference includes:

[0021] A preset mapping relationship between brightness difference and target aperture value is established. Based on the mapping relationship, the target aperture value is determined according to the brightness difference, and the actual aperture value of the aperture stop is adjusted to reach the target aperture value.

[0022] Furthermore, the mapping relationship includes orifice calculation formulas corresponding to different liquid level heights. The orifice calculation formulas are determined based on the actual liquid level heights, and the target orifice value is determined based on the corresponding orifice calculation formulas and the brightness difference.

[0023] As can be seen from the above technical solutions, the embodiments of this application have at least the following beneficial effects: by first completing the whole-hole imaging with a preset aperture value using an aperture stop, the brightness difference between the center position and other different positions inside the hole is obtained, and the actual aperture value of the aperture stop is adjusted according to the brightness difference during the next imaging, so that other different positions, such as the edge area, can have large-angle light and the concave liquid surface shape to form a cancellation effect, solving the problem of edge blackening caused by the concave liquid surface formed by the surface tension of the liquid, directly improving the brightness of the edge area of ​​the concave liquid surface, thereby reducing cell loss in the edge area during imaging; at the same time, this method does not require a preset fixed compensation mode, and the aperture stop can dynamically adapt to various concave liquid surface morphologies caused by changes in sample volume, ultimately obtaining a high-quality image of the whole hole while ensuring imaging resolution and material compatibility.

[0024] A second aspect of this application provides an imaging device, including: an imaging module, an illumination module, and a control module;

[0025] The lighting module includes an aperture stop with adjustable aperture.

[0026] The imaging module and the illumination module are configured to acquire images.

[0027] The control module is used to issue control commands, and the imaging module and the illumination module are configured to respond to the control commands respectively to implement the imaging method as described in the first aspect embodiment of this application.

[0028] Furthermore, the aperture stop is adjustable through stepless adjustment or gear shifting adjustment.

[0029] A third aspect of this application provides an electronic device, including: a memory, a processor, and a computer program stored in the memory and executable on the processor, wherein the processor executes the computer program to implement the imaging method as described in the first aspect of this application.

[0030] The storage medium of the fourth aspect of this application stores a computer program thereon, which, when executed by a processor, implements the imaging method as described in the first aspect of this application.

[0031] It is easy to understand that the imaging device in the second aspect embodiment of this application, the electronic device in the third aspect embodiment of this application, and the storage medium in the fourth aspect embodiment of this application all have the same technical effects as the imaging method in the first aspect embodiment, and therefore will not be described again.

[0032] Additional aspects and advantages of this application will be set forth in part in the description which follows, and in part will be obvious from the description, or may be learned by practice of this application. Attached Figure Description

[0033] To more clearly illustrate the technical solutions in the embodiments of this application or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of this application. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0034] Figure 1 An exploded view provided for one embodiment of this application;

[0035] Figure 2 This is a schematic diagram of the optical path for small-aperture imaging provided in one embodiment of this application;

[0036] Figure 3 This is a schematic diagram of the optical path for large-aperture imaging provided in one embodiment of this application;

[0037] Figure 4 A schematic diagram illustrating the division of the hole positions according to one embodiment of this application;

[0038] Figure 5 This is a system diagram provided for one embodiment of this application.

[0039] Figure label:

[0040] 110. Illumination module; 111. Aperture stop; 120. Imaging module; 130. Perforation plate device; 131. Aperture position; 1311. Central region; 1312. Edge region;

[0041] 201. Processor; 202. Memory. Detailed Implementation

[0042] 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.

[0043] See Figures 1 to 5 As shown, an embodiment of the first aspect of this application discloses an imaging method, including the following steps:

[0044] The aperture stop 111, with a preset aperture value, works in conjunction with the imaging module 120 to acquire a preliminary image of the sample at at least one aperture position 131 in the orifice plate device 130.

[0045] Calculate the brightness difference between the center position and other different positions within aperture 131 based on the preliminary image obtained from the preliminary image acquisition.

[0046] The actual aperture value of aperture stop 111 is adjusted according to the brightness difference to cooperate with imaging module 120 to acquire actual images of the sample at each position within aperture 131.

[0047] In the embodiments of this application, a full-well image is first taken using an aperture stop 111 with a preset aperture value to obtain the brightness difference between the center position and other different positions within the well. During the next image capture, the actual aperture value of the aperture stop 111 is adjusted according to the brightness difference, so that other different positions, such as the edge region 1312, can have large-angle light that cancels out the concave liquid surface shape, solving the problem of edge blackening caused by the concave liquid surface formed by liquid surface tension. This directly improves the brightness of the edge region 1312 of the concave liquid surface, thereby reducing cell loss in the edge region 1312 during imaging. At the same time, this method does not require a preset fixed compensation mode, and the aperture stop 111 can dynamically adapt to various concave liquid surface morphologies caused by changes in sample volume. Ultimately, a high-quality image of the entire well is obtained while ensuring imaging resolution and material compatibility.

[0048] Understandably, the aperture stop 111 is adjustable in size to change the light emission angle of the illumination module 110. The brightness difference between different locations, such as the center and edge of the aperture 131, is primarily caused by the refraction of light by the concave liquid surface formed by the surface tension of the liquid. This manifests as lower brightness and darkening at the edges, leading to image failure and cell loss at these locations. Therefore, by matching the brightness difference to the aperture adjustment, the amount of light entering and the brightness of the edge region 1312 of the aperture 131 are specifically increased. This directly improves the problems of darkening and loss of detail in edge imaging caused by the concave liquid surface, reducing the occurrence of cell loss in the edge region 1312 during imaging.

[0049] It should be noted that this method does not require a preset fixed compensation mode. The adjustment of the aperture stop 111 dynamically changes with the brightness difference, adapting to various concave liquid surface morphologies caused by different sample volumes. It exhibits strong adaptability, meaning there is no need to adjust fixed parameters for different sample volumes. In use, supplementary lighting is achieved simply by adjusting the aperture value of the aperture stop 111, without reducing the numerical aperture of the objective lens or changing the core parameters of the imaging module 120. Furthermore, it requires no modification to existing universal well plate consumables, ensuring compatibility with mainstream industry consumables. This achieves improved overall well imaging quality while effectively ensuring imaging resolution and material compatibility. Moreover, implementing this method for imaging allows for more uniform imaging brightness across the center and edges of the well 131, resulting in clear and complete details, achieving high-quality imaging of the entire area of ​​a single well, and avoiding the problem of imaging failure in localized areas.

[0050] In some embodiments, the imaging module 120 first takes a preliminary picture of the entire area of ​​the aperture position 131 using the preset aperture value of the aperture stop 111 to obtain complete imaging data inside the aperture. Based on the obtained preliminary image, the brightness difference between different positions such as the center and edge of the aperture position 131 is calculated. Then, the actual aperture value of the aperture stop 111 is adjusted according to the brightness difference to allow the edge area 1312 to capture light at a large angle, forming a counteracting effect with the concave liquid surface morphology. Finally, the actual image is acquired at each position inside the aperture. The core logic of the imaging method in this embodiment is to use the brightness difference as the control basis and compensate for the difference in light loss of the concave liquid surface by dynamically adjusting the aperture stop 111.

[0051] The following will combine Figures 1 to 5 The imaging methods disclosed in the embodiments of this application will be explained and described in detail.

[0052] Understandably, after initial image acquisition of the sample, the imaging information is processed by the control module to calculate the brightness difference between the center position and other different positions, which serves as the basis for adjusting the aperture stop 111. In some embodiments, the brightness difference is obtained through unified imaging, with the control module processing the initial image information uniformly. In other embodiments, the brightness difference is obtained by first imaging in zones, and then processing the imaging data of each zone separately through the control module.

[0053] In some specific embodiments, preliminary image acquisition of the sample is performed, including:

[0054] Select the target aperture to be imaged, take multiple images of the target aperture, and fuse the images after the imaging area covers the cross-sectional area of ​​the aperture to obtain a preliminary image.

[0055] Exemplary examples include, in some embodiments, preliminary image acquisition of the sample, including:

[0056] Two-dimensional array image acquisition is performed to obtain multiple sub-images. The acquired sub-images are then fused to obtain a preliminary image of the sample.

[0057] It should be noted that in the imaging method of this embodiment, a complete image of the entire aperture position 131 is obtained by stitching together multiple sub-images, providing a more accurate benchmark for brightness difference calculation. Specifically, the imaging module 120 captures A*B (A>1, B>1) images to complete the two-dimensional array image acquisition step. After obtaining multiple sub-images, they are fused into a complete aperture position map. By covering the entire area of ​​the aperture position 131 with image acquisition, the situation of insufficient field of view in a single acquisition is avoided, ensuring the quality of the initial image.

[0058] Understandably, the accuracy of the brightness information in the fused image is improved, providing a data foundation for subsequent calculations of the brightness difference between the center and edge positions, and ensuring the accuracy of the dynamic adjustment of the aperture stop 111. Based on this, the fused aperture map is divided into central region 1311 and edge region 1312 according to the image brightness values, so as to calculate the brightness difference in the subsequent calculation.

[0059] For example, in some embodiments, calculating the brightness difference between the center position within the aperture 131 and other different positions based on the preliminary image obtained from the preliminary image acquisition includes:

[0060] Based on the image brightness value, the preliminary image obtained by fusion is divided into regions to obtain the central region 1311 and the edge region 1312. The difference between the image brightness value of the central region 1311 and the image brightness value of the edge region 1312 is calculated to obtain the brightness difference value.

[0061] It should be noted that, based on the image brightness, the preliminary image is divided into a central region 1311 and an edge region 1312. The difference is calculated to quantify the degree of edge brightness attenuation caused by the concave liquid surface. After obtaining the quantized brightness difference data that can be used for adjustment, it can provide a basis for the dynamic adjustment of the aperture stop 111. By specifically compensating for the edge brightness, the problem of blackening at the edge of the concave liquid surface can be solved.

[0062] It is understood that the division of the central region 1311 and the edge region 1312 can be based on the field of view of the imaging module 120 and / or the size of the aperture 131, and implemented with a preset division logic. In some embodiments, the division method can be based on experience or experimentation to ensure that the central region 1311 can obtain a high-quality image at the preset aperture value of the aperture stop 111 after division, while other regions, such as the edge region 1312 or the intermediate region between the central region 1311 and the edge region 1312, can achieve high-quality imaging according to their corresponding actual aperture values. In other embodiments, the aperture 131 is divided to form a number of regions. Specifically, the aperture 131 can be matrix-divided equally, with a certain number of regions near the center of the aperture 131 designated as the central region 1311, and the remaining regions as the edge region 1312. This allows for precise differentiation of the concave liquid surface light-loss-free central region 1311, the light-attenuated edge region 1312, and the intermediate regions in between, avoiding brightness data deviations caused by ambiguous partitioning and improving the targeting of subsequent aperture adjustments.

[0063] Exemplary, in some embodiments, reference is made to Figure 4 The hole position 131 is divided into 16 equal regions. The four regions closest to the center of the hole position 131 are designated as the central region 1311, and the remaining regions are designated as the edge region 1312.

[0064] It should be noted that the aperture position 131 is divided into 16 equal parts in a standardized area. The four sections closest to the center of the aperture are designated as the central area 1311, and the remaining 12 sections are designated as the edge area 1312. By defining the areas of light difference on the concave liquid surface, the brightness difference calculation is made to fit the actual light attenuation law of the aperture position 131, providing a quantitative basis for the dynamic adjustment of the aperture stop 111.

[0065] Exemplary, in some embodiments, reference is made to Figure 2 and Figure 3 The imaging module 120, in conjunction with other imaging modules, acquires actual images of the sample at various locations within the aperture 131, including:

[0066] Actual image acquisition is performed based on the central region 1311 and the edge region 1312 respectively;

[0067] When the central region 1311 is actually acquiring images, the aperture stop 111 uses a preset aperture value to cooperate with the imaging module 120 to acquire images.

[0068] When the edge region 1312 is actually acquiring an image, the aperture stop 111 is adjusted to the target brightness value based on the brightness value when the central region 1311 is imaged. After the actual brightness value of the edge region 1312 reaches the target brightness value, the image is acquired in conjunction with the imaging module 120.

[0069] It is understandable that when the aperture stop 111 is used with the imaging module 120 to acquire images with a preset aperture value, it can meet the imaging quality requirements of the central region 1311 under normal conditions. For the edge region 1312, the actual aperture value of the aperture stop 111 is dynamically adjusted based on the target brightness value until the actual imaging brightness of the edge region 1312 is completely matched with the central brightness, so as to avoid the loss of cell imaging in the edge region 1312 to the greatest extent.

[0070] In some embodiments, 12 edge regions 1312 cover all edge regions 1312 of the aperture 131. Based on the aperture adjustment of brightness difference, each edge partition can take in a suitable large-angle light, which can offset the light loss of the concave liquid surface, comprehensively improve the overall brightness of the edge, minimize the loss of cell imaging in the entire edge region 1312 of the aperture 131, and finally achieve high-quality imaging with balanced brightness and complete details in the center and edge of the aperture 131.

[0071] It is understandable that the actual aperture value of the aperture stop 111 is adjusted according to the brightness difference to ensure that the opening size of the aperture stop 111 is adjusted to a suitable size and cooperates with the imaging module 120 when imaging different areas. This ensures that the brightness meets the imaging quality requirements when imaging different areas. In some embodiments, the actual aperture value of the aperture stop 111 can be adjusted through closed-loop control to achieve precise adjustment and meet the imaging quality requirements by gradually eliminating the brightness difference. In other embodiments, the target aperture value can be calculated using the brightness difference as the independent variable, and then the actual aperture value of the aperture stop 111 can be adjusted to the target aperture value to meet the imaging requirements.

[0072] For example, in some embodiments, the actual aperture value of the aperture stop 111 is adjusted according to the brightness difference, including:

[0073] A mapping relationship between the preset brightness difference and the target aperture value is established. Based on the mapping relationship, the target aperture value is determined according to the brightness difference. The actual aperture value of the aperture stop 111 is adjusted to achieve the target aperture value.

[0074] Understandably, the mapping relationship between the brightness difference and the target aperture value provides a clear quantitative basis for aperture adjustment, accurately translating the brightness attenuation caused by the concave surface into the adjustment amount of the aperture stop 111, thus improving the accuracy of edge illumination. Specifically, the precisely matched target aperture value allows the large-angle light captured by the edge region 1312 to offset the light loss from the concave surface, solving problems such as edge darkening. Since the target aperture can be determined by matching the brightness difference mapping relationship, the adjustment time of the aperture stop 111 is significantly shortened, making the overall imaging acquisition process more efficient and adapting to the rapid imaging needs of batch aperture positions 131.

[0075] In some embodiments, the mapping relationship between brightness difference and target aperture value can be constructed based on experience or experimentation. In the mapping relationship between brightness difference and target aperture value, the brightness difference and the target aperture value are mapped one-to-one. The capacity supply range corresponding to the brightness difference can be determined by the mapping relationship between brightness difference and target aperture value.

[0076] Understandably, due to the surface tension of the liquid, a concave meniscus will form within the well plate. The shape and size of this concave meniscus are not fixed; they are directly related to the amount of sample added to the well plate. Different sample amounts will lead to differences in the liquid level, thus causing changes in the shape of the concave meniscus. Therefore, the construction of the mapping relationship needs to take into account the difference in liquid level caused by different sample amounts added to the well plate.

[0077] For example, in some embodiments, the mapping relationship includes orifice calculation formulas corresponding to different liquid level heights. The orifice calculation formula is determined based on the actual liquid level height, and the target orifice value is determined based on the corresponding orifice calculation formula and the brightness difference. It is understood that different liquid level heights correspond to different concave liquid surface morphologies, and the corresponding orifice calculation formulas can be specifically matched to the light loss patterns of each morphology, solving the problem of incomplete adaptation by a single mapping relationship. Orifice sizes can be calculated for various sample loading amounts under common experimental scenarios, thereby covering as many concave liquid surface scenarios as possible.

[0078] It should be noted that the logic of classifying and adapting the calculations by liquid level height uses a formula to convert the two parameters of liquid level height and brightness difference into a precise aperture adjustment amount, so that large-angle light and concave liquid surfaces of different shapes can form a precise cancellation effect, achieving a more refined dynamic adaptation.

[0079] In some embodiments, the partitioned imaging fusion is first completed by using a preset aperture, the brightness difference between the center and the edge is calculated, the actual liquid level in the hole is first identified, the corresponding aperture calculation formula is matched, and then the measured brightness difference is substituted into the matched formula to accurately calculate the target aperture value. The aperture stop 111 is adjusted to this value. During actual imaging, the preset aperture is used in the center, and the target aperture is used to supplement the large-angle light at the edge so that the brightness matches the center and the acquisition is completed.

[0080] In one specific embodiment, the imaging method of this embodiment includes the following steps:

[0081] S1: The aperture stop 111, with a preset aperture value, works with the imaging module 120 to acquire a preliminary image of the sample at least one aperture position 131 in the orifice plate device 130. When acquiring the preliminary image of the sample, a 4*4 two-dimensional array image is acquired at the aperture position 131 to obtain multiple sub-images. The acquired multiple sub-images are fused to obtain a preliminary image of the sample.

[0082] S2: Calculate the brightness difference between the center position and other different positions within the aperture 131 based on the preliminary image obtained from the preliminary image acquisition. Specifically, based on the image brightness value of the fused preliminary image, the preliminary image is divided into regions to obtain the central region 1311 and the edge region 1312. Calculate the difference between the image brightness value of the central region 1311 and the image brightness value of the edge region 1312 to obtain the brightness difference.

[0083] S3: Perform actual image acquisition based on the central region 1311 and the edge region 1312 respectively;

[0084] When the central region 1311 is actually acquiring images, the aperture stop 111 uses a preset aperture value to cooperate with the imaging module 120 to acquire images.

[0085] When the edge region 1312 is actually imaged, the brightness value of the central region 1311 is used as the target brightness value. The corresponding aperture calculation formula is determined according to the actual liquid level height. Based on the corresponding aperture calculation formula and the brightness difference, the target aperture value is determined and the actual aperture value of the aperture stop 111 is adjusted to reach the target aperture value. After the actual brightness value of the edge region 1312 reaches the target brightness value, the image is acquired in conjunction with the imaging module 120.

[0086] The imaging method of this application embodiment is described in detail below with reference to a specific example. It should be noted that the following embodiment is merely an exemplary description and should not be construed as limiting the embodiments of this application.

[0087] See Figures 1 to 5 As shown, the imaging method of this embodiment first performs a full-aperture photograph using the standard aperture stop 111 size. Then, it analyzes and calculates the brightness ratio between the darkened edge region and the central region 1311 to obtain the brightness difference at different locations. Different brightness differences are pre-set to correspond to specific aperture stop 111 opening diameters. Then, a full-aperture photograph is performed again. When photographing the edge region 1312, the size of the motorized aperture stop 111 is adjusted to increase, and when photographing the central region 1311, it is decreased. By changing the opening size of the aperture stop 111 of the illumination system, the numerical aperture of the illumination system is adjusted. The larger the opening of the aperture stop 111, the larger the angle of the light emitted from the illumination system. When the large angle of the edge light passes through the concave surface, it can cancel out the shape of the concave surface, thereby increasing the brightness of the concave surface region.

[0088] Furthermore, different sizes of well plates correspond to different commonly used liquid surface volumes in experiments, and different liquid surface volumes will result in different brightness differences in the liquid surface height. Taking a 96-well plate as an example, commonly used liquid surface volumes are 50µL, 100µL, 150µL, and 200µL. For example, with a 96-well plate and a volume of 50µL, the brightness of the concave liquid surface at the edge is 30% of that in the central region (1311); with 100µL, the brightness is 60% of that in the central region (1311); with 150µL, the brightness is 67% of that in the central region (1311); and with 200µL, the brightness is 70% of that in the central region (1311).

[0089] When imaging the well plate, each well is divided into 16 equal regions for photographing and image fusion. The four images of the central region are designated as the central region 1311, and the twelve images of the outer region are designated as the edge region 1312. First, the aperture stop 111 opening size is kept constant at 8mm for photographing. The brightness values ​​of the edge region 1312 and the central region 1311 are obtained by calculating the brightness values ​​of the images, with the brightness value of the central region 1311 being the target value. The opening size of the aperture stop 111 is calculated using a formula, thus obtaining the aperture stop 111 opening size for the edge region 1312. Since liquids absorb light, different liquid volumes will affect the brightness value, and different culture media also have different degrees of light absorption. First, the RGB values ​​of the colors inside the well are photographed, and then matched with pre-photographed images of various culture media using an algorithm to obtain the correlation parameters between liquid volume and brightness value for that culture media type, as well as the correlation parameters between the target brightness value, liquid volume, and aperture stop. Then, based on the brightness value of the central region 1311 and the correlation parameter between the liquid surface volume and the brightness value (the liquid surface volume and the brightness value show a linear decreasing trend), the liquid surface volume at this time is calculated (due to the existence of liquid evaporation, there may be a situation where the actual liquid surface volume is smaller than the liquid surface volume when the sample is added). The target brightness value y is linearly related to the aperture stop x, and the target brightness value y = kx + b, where the values ​​of k and b are linked to the actual liquid surface volume v. Based on the target brightness value, the liquid surface volume and the correlation parameter between the aperture stop, the opening size of the aperture stop 111 is calculated. Then, a second photo is taken. When taking a picture of the edge position, the opening size of the aperture stop 111 is changed, thereby improving the brightness value of the edge region 1312 and making the whole aperture imaging effect better. For example, in this system, the brightness value of the central region of commonly used transparent liquids such as PBS can be obtained as y1 = -0.112v + 57.5 based on the correlation parameters between the liquid surface volume and brightness value under this culture medium type. The actual liquid surface volume v can be obtained through this formula. Based on the target brightness value, liquid surface volume, and aperture stop correlation parameters under this culture medium type, the k and b values ​​of commonly used transparent liquids such as PBS are correlated with the actual liquid surface volume v in this system as follows: k = 0.5349v + 2.7801, b = -4.3253v + 10.968. Therefore, based on the target brightness value y = kx + b, the opening size of the aperture stop x can be calculated. In other embodiments, based on the above imaging logic, the coefficients and / or constants used in the above calculation process can be adaptively adjusted according to data obtained from actual experiments or based on empirical data to meet the needs of different application scenarios.

[0090] It is understandable that the imaging method of this embodiment solves the problem of the edge region 1312 light being refracted and unable to be received due to the concave liquid surface by first initially acquiring the image and then automatically calculating and completing the imaging of different regions with different numerical apertures of the matching illumination system. Furthermore, this method can adapt to different well plates and different liquid filling heights in the well plates, so that when performing whole-hole imaging on well plates with concave liquid surfaces, it can obtain high-quality images with better illumination at the edge concave liquid surface and reduce the problem of 1312 cell loss in the edge region.

[0091] An embodiment of the second aspect of this application discloses an imaging device with an aperture stop 111, specifically referring to... Figure 1 It includes: an imaging module 120, an illumination module 110, and a control module;

[0092] The illumination module 110 includes an aperture stop 111 with an adjustable aperture; the imaging module 120 is configured to cooperate with the illumination module 110 to acquire images; the control module is used to issue control commands, and the imaging module 120 and the illumination module 110 are configured to respond to the control commands respectively to implement the imaging method disclosed in the first aspect of the present application.

[0093] In some embodiments, the aperture stop 111 is adjustable through stepless adjustment or step-by-step adjustment. It is understood that an adjustable aperture stop 111 is designed in the illumination module 110 to change the light emission angle of the illumination module 110, thereby solving the blackening problem caused by the concave liquid surface in the imaging.

[0094] In some embodiments, the infinitely adjustable aperture stop 111 can be an electric aperture stop 111 or a mechanical blade aperture stop 111, etc., so that different sizes of openings can be adjusted. The aperture stop 111 with shift adjustment can be selected from multiple aperture plates including fixed openings, and the opening size of the aperture stop 111 can be adjusted by switching through a motor.

[0095] It is understood that the content of the above method embodiments is applicable to the present device embodiments. The specific functions implemented by the present device embodiments are the same as those of the above method embodiments, and the beneficial effects achieved are also the same as those achieved by the above method embodiments.

[0096] Reference Figure 5 An electronic device according to a third aspect of this application includes: a memory 202, a processor 201, and a computer program stored on the memory 202 and executable on the processor 201. When the processor 201 executes the computer program, it implements the method embodiment described above.

[0097] Similarly, it can be understood that the content of the above method embodiments is applicable to this device embodiment. The specific functions implemented by this device embodiment are the same as those of the above method embodiments, and the beneficial effects achieved are also the same as those achieved by the above method embodiments.

[0098] The storage medium of the fourth aspect of this application stores a computer program thereon, which, when executed by a processor, implements the imaging method of the first aspect of this application.

[0099] Similarly, the content of the above method embodiments is applicable to the present computer-readable storage medium embodiments. The specific functions implemented by the present computer-readable storage medium embodiments are the same as those of the above method embodiments, and the beneficial effects achieved are also the same as those achieved by the above method embodiments.

[0100] In the description of this application, it should be understood that the terms "center", "upper", "lower", "front", "rear", "left", "right", "vertical", "horizontal", "top", "bottom", "inner", "outer", etc., indicate the orientation or positional relationship based on the orientation or positional relationship shown in the accompanying drawings. They are only for the convenience of describing this application and simplifying the description, and do not indicate or imply that the device or element referred to must have a specific orientation, or be constructed and operated in a specific orientation. Therefore, they should not be construed as limitations on this application.

[0101] The terms "first" and "second" are used for descriptive purposes only and should not be construed as indicating or implying relative importance or implicitly specifying the number of technical features indicated. Therefore, a feature defined as "first" or "second" may explicitly or implicitly include one or more of that feature. In the description of this application, unless otherwise stated, "multiple" means two or more. It should be noted that the term "and / or" used herein is merely a description of the relationship between related objects, indicating that three relationships can exist. For example, A and / or B can represent: A alone, A and B simultaneously, or B alone. Similarly, at least one of A or B can also represent: A alone, A and B simultaneously, or B alone.

[0102] In the description of this application, it should be noted that, unless otherwise expressly specified and limited, the terms "installation," "connection," and "joining" should be interpreted broadly. For example, they can refer to a fixed connection, a detachable connection, or an integral connection; they can refer to a direct connection or an indirect connection through an intermediate medium; and they can refer to the internal connection between two components. Those skilled in the art can understand the specific meaning of the above terms in this application based on the specific circumstances.

[0103] In the foregoing description of this specification, references to terms such as "one embodiment," "another embodiment," or "some embodiments" indicate that a specific feature, structure, material, or characteristic described in connection with an embodiment or example is included in at least one embodiment or example of this application. In the description of this specification, specific features, structures, materials, or characteristics may be combined in any suitable manner in one or more embodiments or examples.

[0104] It should be understood that although the steps in the flowcharts of the accompanying figures are shown sequentially as indicated by the arrows, these steps are not necessarily executed in the order indicated by the arrows. Unless explicitly stated herein, there is no strict order restriction on the execution of these steps, and they can be executed in other orders. Moreover, at least some steps in the flowcharts of the accompanying figures may include multiple sub-steps or multiple stages. These sub-steps or stages are not necessarily completed at the same time, but can be executed at different times, and their execution order is not necessarily sequential, but can be performed alternately or in turn with other steps or at least some of the sub-steps or stages of other steps.

[0105] Furthermore, although this application is described in the context of functional modules, it should be understood that, unless otherwise stated to the contrary, one or more of the functions and / or features may be integrated into a single physical device and / or software module, or one or more functions and / or features may be implemented in a separate physical device or software module. It is also understood that a detailed discussion of the actual implementation of each module is unnecessary for understanding this application. Rather, given the properties, functions, and internal relationships of the various functional modules in the apparatus disclosed herein, the actual implementation of the module will be understood within the scope of conventional technology for an engineer. Therefore, those skilled in the art can implement the application set forth in the claims using ordinary techniques without excessive experimentation. It is also understood that the specific concepts disclosed are merely illustrative and not intended to limit the scope of this application, which is determined by the full scope of the appended claims and their equivalents.

[0106] If a function is implemented as a software functional unit and sold or used as an independent product, it can be stored in a computer-readable storage medium. Based on this understanding, the technical solution of this application, in essence, or the part that contributes to the prior art, or a part of the technical solution, can be embodied in the form of a software product. This computer software product is stored in a storage medium and includes several instructions to cause a computer device (which may be a personal computer, server, or network device, etc.) to execute all or part of the steps of the methods of the various embodiments of this application. The aforementioned storage medium includes various media capable of storing program code, such as USB flash drives, portable hard drives, read-only memory (ROM), random access memory (RAM), magnetic disks, or optical disks.

[0107] The logic and / or steps represented in the flowchart or otherwise described herein, for example, can be considered as a sequenced list of executable instructions for implementing logical functions, and can be embodied in any computer-readable medium for use by, or in conjunction with, an instruction execution system, apparatus, or device (such as a computer-based system, a processor-included system, or other system that can fetch and execute instructions from, an instruction execution system, apparatus, or device). For the purposes of this specification, "computer-readable medium" can be any means that can contain, store, communicate, propagate, or transmit programs for use by, or in conjunction with, an instruction execution system, apparatus, or device.

[0108] More specific examples of computer-readable media (a non-exhaustive list) include: electrical connections (electronic devices) having one or more wires, portable computer disk drives (magnetic devices), random access memory (RAM), read-only memory (ROM), erasable and editable read-only memory (EPROM or flash memory), fiber optic devices, and portable optical disc read-only memory (CDROM). Furthermore, computer-readable media can even be paper or other suitable media on which programs can be printed, because programs can be obtained electronically, for example, by optically scanning the paper or other medium, followed by editing, interpreting, or otherwise processing as necessary, and then stored in computer memory.

[0109] It should be understood that various parts of this application can be implemented using hardware, software, firmware, or a combination thereof. In the above embodiments, multiple steps or methods can be implemented using software or firmware stored in memory and executed by a suitable instruction execution system. For example, if implemented in hardware, as in another embodiment, it can be implemented using any one or a combination of the following techniques known in the art: discrete logic circuits having logic gates for implementing logical functions on data signals, application-specific integrated circuits (ASICs) having suitable combinational logic gates, programmable gate arrays (PGAs), field-programmable gate arrays (FPGAs), etc.

Claims

1. An imaging method, characterized in that, Includes the following steps: The aperture stop, with a preset aperture value, works in conjunction with the imaging module to acquire a preliminary image of the sample at at least one well in the orifice plate device. Calculate the brightness difference between the center position and other different positions within the aperture based on the preliminary image obtained from the preliminary image acquisition. The actual aperture value of the aperture stop is adjusted according to the brightness difference to cooperate with the imaging module to acquire actual images of the sample at each position within the aperture.

2. The imaging method according to claim 1, characterized in that: The preliminary image acquisition of the sample includes: Select the target aperture to be imaged, take multiple images of the target aperture, and fuse the images after the imaging area covers the cross-sectional area of ​​the aperture to obtain the preliminary image.

3. The imaging method according to claim 2, characterized in that: The step of calculating the brightness difference between the center position within the aperture and other different positions based on the preliminary image obtained from the preliminary image acquisition includes: Based on the image brightness value, the preliminary image obtained by fusion is divided into regions to obtain a central region and an edge region. The difference between the image brightness value of the central region and the image brightness value of the edge region is calculated to obtain the brightness difference value.

4. The imaging method according to claim 3, characterized in that: The process of acquiring actual images of the sample at each location within the aperture, in conjunction with the imaging module, includes: Actual image acquisition is performed based on the central region and the edge region respectively; When the central region is actually image acquired, the aperture stop cooperates with the imaging module to acquire the image using the preset aperture value. When the edge region is actually captured, the aperture stop is adjusted to the target brightness value based on the brightness value of the central region during imaging. After the actual brightness value of the edge region reaches the target brightness value, the image is captured in conjunction with the imaging module.

5. The imaging method according to claim 1, characterized in that: The step of adjusting the actual aperture value of the aperture stop according to the brightness difference includes: A preset mapping relationship between brightness difference and target aperture value is established. Based on the mapping relationship, the target aperture value is determined according to the brightness difference, and the actual aperture value of the aperture stop is adjusted to reach the target aperture value.

6. The imaging method according to claim 5, characterized in that: The mapping relationship includes orifice calculation formulas corresponding to different liquid level heights. The orifice calculation formulas are determined based on the actual liquid level heights. The target orifice value is determined based on the corresponding orifice calculation formulas and the brightness difference.

7. An imaging device, characterized in that, include: Imaging module, illumination module, and control module; The lighting module includes an aperture stop with adjustable aperture. The imaging module and the illumination module are configured to acquire images. The control module is used to issue control commands, and the imaging module and the illumination module are configured to respond to the control commands respectively to implement the imaging method as described in any one of claims 1 to 6.

8. The imaging apparatus according to claim 7, characterized in that: The aperture stop is adjustable through stepless adjustment or gear shifting adjustment.

9. An electronic device, comprising: A memory, a processor, and a computer program stored in the memory and executable on the processor, characterized in that the processor, when executing the computer program, implements the imaging method as described in any one of claims 1 to 6.

10. A storage medium having a computer program stored thereon, characterized in that, When the computer program is executed by a processor, it implements the imaging method as described in any one of claims 1 to 6.

Images