CN-116363095-B - Methods, devices, apparatus and media for determining phagocytic capacity of phagocytes

Abstract

A method for determining phagocytic capacity of a phagocyte includes obtaining a phagocytic image and a target cell image extracted from a high-content image obtained by photographing co-cultured phagocytes and target cells, decomposing the target cell image into a plurality of sub-band images each including pixels of the target cell image having a luminance value falling in a corresponding one of a plurality of luminance sub-bands obtained by dividing a luminance range of the pixels of the target cell image, determining respective numbers of target cell objects respectively included in the plurality of sub-band images, determining the numbers of phagocytes included in the phagocytic image, and determining the phagocytic capacity of the phagocytes to the target cells based on the respective numbers of target cell objects respectively included in the plurality of sub-band images and the numbers of phagocytes included in the phagocytic image.

Inventors

• WANG CHENLONG
• ZHANG YILAN
• YANG CHANGMEI
• CHEN QIANQIAN
• SONG LE

Assignees

• 百图生科(北京)智能技术有限公司

Dates

Publication Date

20260512

Application Date

20230328

Priority Date

20221219

Claims (20)

1. 1. A method for determining phagocytic capacity of phagocytes, comprising: The method comprises the steps of obtaining a target cell image, wherein the target cell image is extracted from a high content image obtained by shooting co-cultured phagocytes and target cells, and the fluorescence intensity of the target cells changes after the target cells are phagocytosed by the phagocytes; Acquiring a phagocyte image containing a plurality of phagocytes, which is extracted from a high content image obtained by photographing co-cultured phagocytes and target cells, and determining the number of phagocytes contained in the phagocyte image; decomposing the target cell image into a plurality of sub-band images, each sub-band image comprising pixels of the target cell image having luminance values falling into a respective one of a plurality of luminance sub-bands; determining the respective numbers of target cell objects contained in the respective plurality of sub-band images, the plurality of luminance sub-bands being partitioned according to fluorescence intensity thresholds, a first one of the fluorescence intensity thresholds being used to distinguish phagocytosed target cells from non-phagocytosed target cells, and Determining phagocytic capacity of the phagocytic cell for the target cell based on the number of target cells falling into at least one of a plurality of luminance subbands, comprising: Calculating a ratio of a weighted sum of the respective numbers of target cell objects contained in the plurality of sub-band images to the number of phagocytes, wherein the ratio indicates phagocytic capacity of the phagocytes to the target cells, and a weight coefficient corresponding to the respective numbers of target cell objects contained in the plurality of sub-band images is positively correlated with a brightness range of the respective brightness sub-bands.
2. 2. The method of claim 1, wherein the method further comprises: Acquiring a blank control group image, wherein the blank control group image is extracted from a high content image obtained by shooting target cells which are independently cultured; and determining the first fluorescence intensity threshold according to the fluorescence intensity of the target cells in the blank control group image.
3. 3. The method of claim 1 or 2, wherein the determining phagocytic capacity of the phagocytic cell for the target cell based on the number of target cells falling into at least one of a plurality of luminance subbands further comprises: determining the phagocytic capacity of the phagocytic cell for the target cell based on the number of target cells falling into at least one of the plurality of luminance subbands and the number of phagocytic cells.
4. 4. The method of claim 1, wherein the determining the respective number of target cell objects respectively contained by the plurality of sub-band images comprises: Graying processing is carried out on the plurality of sub-band images; And carrying out image segmentation on the sub-band images in the plurality of sub-band images after graying based on an adaptive threshold algorithm to obtain a first image containing a first number of first sub-areas, wherein the first sub-areas indicate target cell areas, and the first number is the number of target cell objects contained in the sub-band images.
5. 5. The method of claim 4, wherein the determining the number of the plurality of target cell objects corresponding to the plurality of subband images, respectively, further comprises: Dividing the first image based on a watershed algorithm to obtain a second image containing a second number of first sub-areas, wherein the second number is greater than or equal to the first number, and the second number is the number of target cell objects contained in the sub-band image.
6. 6. The method of claim 5, wherein the determining the number of the plurality of target cell objects corresponding to the plurality of subband images, respectively, further comprises: filtering out at least one first sub-region of the second number of first sub-regions to obtain a third number of first sub-regions, wherein the area of each of the at least one first sub-region is smaller than a first area threshold and/or the area of each of the at least one first sub-region is larger than a second area threshold, and the third number is the number of target cell objects contained in the sub-band image.
7. 7. The method of claim 1, wherein the determining the number of phagocytes contained in the phagocyte image comprises: Graying treatment is carried out on the phagocyte image; And performing image segmentation on the phagocyte image after graying based on an adaptive threshold algorithm to obtain a fourth image containing a fourth number of second subregions, wherein the second subregions indicate phagocyte regions, and the fourth number is the number of phagocytes contained in the phagocyte image.
8. 8. The method of claim 7, wherein the determining the number of phagocytes contained in the phagocyte image further comprises: And image segmentation is carried out on the fourth image based on a watershed algorithm to obtain a fifth image containing a fifth number of subareas, wherein the fifth number is larger than or equal to the fourth number, and the fifth number is the number of phagocytes contained in the phagocyte image.
9. 9. The method of claim 8, wherein the determining the number of phagocytes contained in the phagocyte image further comprises: Filtering out at least one second subregion of the fifth number of second subregions to obtain a sixth number of second subregions, wherein the area of each of the at least one second subregion is smaller than a third area threshold and/or the area of each of the at least one second subregion is larger than a fourth area threshold.
10. 10. The method of claim 1, wherein the acquiring the image of the target cell comprises: acquiring a high content image shot by a high content cell imaging system on co-cultured phagocytes and target cells, wherein the high content image is provided with a second color channel, and the second color channel corresponds to the target cell object; Preprocessing the high content image to generate a color image; And generating the target cell image according to the image information corresponding to the second color channel of the color image.
11. 11. The method of claim 1, further comprising: Prior to said determining the number of phagocytes contained in said phagocyte image: obtaining a denoising image, wherein the denoising image has the same size as the phagocyte image, and pixels in a middle region of the denoising image have pixel values for compensating for halation noise; And subtracting the pixel value of each pixel of the denoising image, which is positioned at the same relative position with each pixel of the phagocyte image, from the pixel value of each pixel of the phagocyte image to obtain the phagocyte image after the vignetting removal.
12. 12. The method of claim 1, wherein the target cell comprises a tumor cell.
13. 13. A method for determining the enhancement or inhibition of phagocytic capacity of phagocytes by a perturbing condition to be tested, comprising: The method according to any one of claims 1 to 12, wherein the target cell image corresponding to the disturbance condition to be tested is extracted from a high content image obtained by photographing co-cultured phagocytes and target cells subjected to the disturbance condition to be tested, the disturbance condition to be tested comprising at least one of exposure to a predetermined concentration of a compound for a predetermined duration, gene editing of candidate genes in the phagocytes and/or the target cells; and determining the enhancement or inhibition effect of the disturbance condition to be tested on the phagocytic capacity of the phagocytes according to the phagocytic capacity of the phagocytes under the disturbance condition to be tested.
14. 14. The method of claim 13, wherein said determining the enhancement or inhibition of phagocytic capacity of phagocytes by said perturbation condition to be tested based on the phagocytic capacity of phagocytes under said perturbation condition to be tested comprises: Determining phagocytic capacity of phagocytes under negative control conditions, wherein the target cell image corresponding to the negative control conditions is extracted from a high content image obtained by photographing the phagocytes and the target cells co-cultured under the negative control conditions, the negative control conditions including conditions that have not been subjected to perturbation or conditions that have been determined to have no effect on phagocytic capacity of the phagocytes; and comparing the phagocytic capacity of the phagocyte under the disturbance condition to be tested with the phagocytic capacity of the phagocyte under the negative control condition, and determining the enhancement or inhibition of the phagocytic capacity of the phagocyte by the disturbance condition to be tested.
15. 15. The method of claim 13, wherein the gene editing comprises gene knockout, gene overexpression, or gene knockdown.
16. 16. An apparatus for determining phagocytic capacity of phagocytes, comprising: The device comprises a first acquisition module, a first acquisition module and a second acquisition module, wherein the first acquisition module is used for acquiring a target cell image, the target cell image is extracted from a high content image obtained by shooting co-cultured phagocytes and target cells, and the fluorescent intensity of the target cells changes after the target cells are phagocytosed by the phagocytes; The device comprises a first acquisition module, a second acquisition module and a control module, wherein the first acquisition module is used for acquiring a phagocyte image and determining the number of phagocytes contained in the phagocyte image; A decomposition module for decomposing the target cell image into a plurality of sub-band images, each sub-band image comprising pixels of the target cell image having luminance values falling into a respective one of a plurality of luminance sub-bands; The device comprises a first determination module, a first judgment module and a second determination module, wherein the first determination module is used for determining the corresponding number of target cell objects respectively contained in the plurality of sub-band images, the plurality of brightness sub-bands are obtained by dividing according to fluorescence intensity thresholds, and a first fluorescence intensity threshold in the fluorescence intensity thresholds is used for distinguishing phagocytosed target cells from non-phagocytosed target cells; a second determining module for determining phagocytic capacity of the phagocytic cell to the target cell based on the number of target cells falling into at least one of the plurality of luminance subbands, comprising: Calculating a ratio of a weighted sum of the respective numbers of target cell objects contained in the plurality of sub-band images to the number of phagocytes, wherein the ratio indicates phagocytic capacity of the phagocytes to the target cells, and a weight coefficient corresponding to the respective numbers of target cell objects contained in the plurality of sub-band images is positively correlated with a brightness range of the respective brightness sub-bands.
17. 17. An apparatus for determining the enhancement or inhibition of phagocytic capacity of phagocytes by a perturbing condition, comprising: a third determination module for determining phagocytic capacity of phagocytes under a disturbance to be tested according to the method of any one of claims 1 to 12, wherein the target cell image corresponding to the disturbance to be tested is extracted from a high content image obtained by photographing co-cultured phagocytes and target cells subjected to the disturbance to be tested, the disturbance to be tested comprising at least one of exposure to a predetermined concentration of a compound for a predetermined duration, gene editing of candidate genes in the phagocytes and/or the target cells; And a fourth determining module, configured to determine, according to the phagocytic capacity of the phagocyte under the disturbance condition to be tested, an enhancement or inhibition effect of the disturbance condition to be tested on the phagocytic capacity of the phagocyte.
18. 18. An electronic device, comprising: Processor, and A memory storing instructions executable by the processor, which when executed by the processor, cause the processor to perform the method of any one of claims 1-15.
19. 19. A non-transitory computer readable storage medium storing instructions which, when executed by a processor, cause the processor to perform the method of any one of claims 1-15.
20. 20. A computer program product comprising instructions, wherein the instructions, when executed by a processor, cause the processor to perform the method of any one of claims 1-15.

Description

Methods, devices, apparatus and media for determining phagocytic capacity of phagocytes The present disclosure claims priority to chinese patent application entitled "method, apparatus, device and medium for determining phagocytic capacity of phagocytes" filed on day 19 12 of 2022 to the national intellectual property agency of the people's republic of China, application number 202211634789.7, the entire contents of which are incorporated herein by reference. Technical Field The present disclosure relates to the field of cell activation state assays, and in particular to a method, apparatus, electronic device, computer readable storage medium and computer program product for determining phagocytic capacity of phagocytes. Background Flow Cytometry (FC) Gate-on/Gate-off technology, determining whether tumor cells phagocytosed by macrophages are positive cells (positive cells indicate that tumor cells are present in macrophages) by an output value (Fluorescence brightness is greater than a preset threshold) of Fluorescence activated cell sorting (Fluorescence ACTIVATED CELL Sorter, FACS), counting the number of positive cells, and determining phagocytic capacity of macrophages under the action of a drug. The greater the number of positive cells, the greater the phagocytic capacity of the macrophages. However, the determination of the positive expression level of positive cells is lacking, and quantitative analysis cannot be achieved. In addition, flow cytometry is required to conduct a number of biological experiments to assess the phagocytic capacity of macrophages. The flux of the flow cytometry instrument is low, and a large amount of manpower and material resources are consumed, so that large-scale biological experiment verification cannot be performed in the task of screening the medicines, and the experimental scale and efficiency of medicine screening are affected. Disclosure of Invention It would be advantageous to provide a mechanism that alleviates, mitigates or even eliminates one or more of the above problems. According to an aspect of the present disclosure, there is provided a method for determining phagocytic capacity of a phagocytic cell, including obtaining a phagocytic cell image and a target cell image, wherein the phagocytic cell image includes a plurality of phagocytic cells, the target cell image includes a plurality of target cell objects, the phagocytic cell image and the target cell image are extracted from a high content image obtained by photographing co-cultured phagocytic cells and target cells, decomposing the target cell image into a plurality of sub-band images, each of which includes a pixel of the target cell image having a luminance value falling into one of a corresponding luminance sub-band of the plurality of luminance sub-bands, the plurality of luminance sub-bands being obtained by dividing a luminance range of the pixel of the target cell image, determining a corresponding number of the target cell objects respectively included in the plurality of sub-band images, determining a number of phagocytic cells included in the target cell image, and determining the phagocytic capacity of the phagocytic cell to the target cell based on the corresponding number of the target cell objects respectively included in the plurality of sub-band images and the number of phagocytic cells included in the phagocytic cell image. According to another aspect of the present disclosure, there is provided a method for determining the enhancement or inhibition of phagocytic capacity of phagocytes by a perturbing condition, comprising acquiring a phagocyte image and a target cell image, wherein the phagocyte image comprises a plurality of phagocytes, the target cell image comprises a plurality of target cell objects, the phagocyte image and the target cell image are extracted from a high content image obtained by photographing co-cultured phagocytes and target cells subjected to the perturbing condition, the perturbing condition comprises at least one of exposing to a compound at a predetermined concentration for a predetermined duration, genetically editing candidate genes in the phagocytes and/or the target cells, decomposing the target cell image into a plurality of subband images, each subband image comprising pixels of the target cell image having a luminance value falling in a corresponding luminance subband of the plurality of luminance subbands, the plurality of luminance subbands being obtained by dividing a luminance range of pixels of the target cell image, determining a corresponding number of target cell objects respectively comprised by the plurality of subband images, determining the number of phagocytes comprised by the phagocyte image, and determining the enhancement of phagocyte capacity of the phagocytes or the target cells comprised by the target cell image based on the corresponding number of target cell objects comprised by the plurality of subband images. According to anothe