Transfection efficiency computing apparatus and method, and microscope and computer-readable storage medium

Abstract

Provided are a transfection efficiency computing apparatus and method, and a microscope and a computer-readable storage medium. The transfection efficiency computing apparatus is used for: acquiring a phase contrast image and a fluorescence image of a cell sample; identifying cells from the phase contrast image, and computing the total number of cells; determining a preset brightness threshold of the fluorescence image, and performing filtering to obtain, from the fluorescence image, effective fluorescence signals with the brightness greater than or equal to the preset brightness threshold; comparing the phase contrast image with the fluorescence image to identify transfection cells, wherein the transfection cells are cells corresponding to the effective fluorescence signals in terms of positions; and on the basis of the total number of cells and the number of transfection cells, computing a transfection efficiency. The provided transfection efficiency computing apparatus can accurately identify the total number of cells and the number of transfection cells, so as to accurately compute a transfection efficiency.

Description

Transfection efficiency calculation device, method, microscope and computer readable storage medium TECHNICAL FIELD

[0001] The present disclosure relates to the technical field of cell transfection, and in particular to a transfection efficiency calculation device, method, microscope and computer readable storage medium. BACKGROUND

[0002] In related art, the ways to calculate the transfection efficiency (transfection success rate) of cells usually include using traditional image processing algorithms and pure artificial intelligence solutions. SUMMARY

[0003] The present inventors have found that the known artificial intelligence solutions in related art are not sensitive to fluorescent signals, and traditional image processing algorithms cannot accurately analyze phase contrast images due to their inherent limitations, so neither of the two methods can accurately calculate the transfection efficiency.

[0004] The present disclosure discloses a transfection efficiency calculation device, method, equipment and computer readable storage medium, which can accurately identify the total number of cells and the number of transfected cells, and then accurately calculate the transfection efficiency.

[0005] The first aspect of the present disclosure provides a transfection efficiency calculation device, which is configured to: acquire a phase contrast image and a fluorescent image of a cell sample; identify cells in the phase contrast image and calculate the total number of cells; determine a preset brightness threshold of the fluorescent image, and filter out effective fluorescent signals in the fluorescent image whose brightness is greater than or equal to the preset brightness threshold; compare the phase contrast image and the fluorescent image to identify transfected cells, the transfected cells being cells corresponding to the positions of the effective fluorescent signals; and calculate the transfection efficiency according to the total number of cells and the number of transfected cells.

[0006] Optionally, the transfected cells are cells corresponding to the positions of the fluorescent regions where the effective fluorescent signals are located, and the area of the fluorescent region corresponding to each transfected cell is greater than or equal to a preset area threshold.

[0007] Optionally, the device is configured to: identify cells in the phase contrast image, mark the outer contour of the cells using a closed contour line, and / or mark the cells using a cell layer.

[0008] Optionally, the device is configured to: superimpose the phase contrast image and the fluorescent image to obtain a superimposed image, the superimposed image including the contour line and / or the cell layer.

[0009] Optionally, the device is configured to determine a feature threshold of the cells, and use the artificial intelligence deep learning capability to identify the cells meeting a feature condition in the phase contrast image according to the feature threshold, the feature condition being that a corresponding feature value of the cell is greater than or equal to the feature threshold, or the feature condition being that the corresponding feature value of the cell is less than or equal to the feature threshold, the feature value being at least one of a cell diameter, a cell radius, and a cell area.

[0010] Optionally, the device is configured to determine the preset brightness threshold according to a user operation, and / or determine the preset brightness threshold according to a fluorescence signal in the fluorescence image.

[0011] Optionally, the device is configured to mark a fluorescence region where the effective fluorescence signal is located using a first color, the first color being different from a second color of other fluorescence regions.

[0012] Optionally, the brightness of the fluorescence region marked using the first color is equal and greater than or equal to the preset brightness threshold.

[0013] Optionally, the device is configured to determine whether the identified transfected cells are accurate, and if it is determined that the identified transfected cells are not accurate, then: redetermine the preset brightness threshold of the fluorescence image; re-compare the phase contrast image and the fluorescence image to identify transfected cells; and if it is determined that the identified transfected cells are accurate, then: calculate the transfection efficiency according to the total number of cells and the number of transfected cells.

[0014] Optionally, the device is configured to perform at least one of the following: redetermine the preset brightness threshold according to a part of the effective fluorescence signal not corresponding to a position of a cell identified in the phase contrast image; and redetermine the preset brightness threshold according to a non-effective fluorescence signal in the fluorescence image having a brightness less than the preset brightness threshold corresponding to at least a part of a position of a cell in the phase contrast image.

[0015] A second aspect embodiment of the present disclosure provides a transfection efficiency calculation method, the method comprising: obtaining a phase contrast image and a fluorescence image of a cell sample; identifying cells in the phase contrast image and calculating a total number of cells; determining a preset brightness threshold of the fluorescence image, and filtering out effective fluorescence signals in the fluorescence image having a brightness greater than or equal to the preset brightness threshold; comparing the phase contrast image and the fluorescence image to identify transfected cells, the transfected cells being cells corresponding to positions of the effective fluorescence signals; and calculating the transfection efficiency according to the total number of cells and the number of transfected cells.

[0016] Optionally, the transfected cells are cells corresponding to positions of the fluorescent regions where the effective fluorescent signals are located, and an area of a corresponding fluorescent region of each of the transfected cells is greater than or equal to a preset area threshold.

[0017] Optionally, the identifying the cells in the phase contrast image and calculating the total number of cells comprises: identifying the cells in the phase contrast image, marking an outer contour of each of the cells with a closed contour line, and / or marking the cells with a cell layer.

[0018] Optionally, the method further comprises: superimposing the phase contrast image and the fluorescent image to obtain a superimposed image, and displaying or not displaying the contour line and / or the cell layer on the superimposed image.

[0019] Optionally, the identifying the cells in the phase contrast image and calculating the total number of cells comprises: determining a feature threshold of the cells, and identifying the cells in the phase contrast image that meet a feature condition according to the feature threshold and an artificial intelligence deep learning capability, the feature condition being that a corresponding feature value of the cells is greater than or equal to the feature threshold or the corresponding feature value of the cells is less than or equal to the feature threshold, the feature value being at least one of a cell diameter, a cell radius, and a cell area.

[0020] Optionally, the determining the preset brightness threshold of the fluorescent image comprises: determining the preset brightness threshold according to a user operation, and / or determining the preset brightness threshold according to a fluorescent signal in the fluorescent image.

[0021] Optionally, the determining the preset brightness threshold of the fluorescent image and filtering out the effective fluorescent signal in the fluorescent image whose brightness is greater than or equal to the preset brightness threshold further comprises: marking a fluorescent region where the effective fluorescent signal is located with a first color, the first color being different from a second color of other fluorescent regions.

[0022] Optionally, the fluorescent regions marked with the first color have equal brightness and the brightness is greater than or equal to the preset brightness threshold.

[0023] Optionally, after the transfected cells are identified, the method further comprises: determining whether the identified transfected cells are accurate, and if it is determined that the identified transfected cells are not accurate, then: re-determining the preset brightness threshold of the fluorescent image, re-comparing the phase contrast image and the fluorescent image to identify the transfected cells, and if it is determined that the identified transfected cells are accurate, then: calculating the transfection efficiency according to the total number of cells and the number of the transfected cells.

[0024] Optionally, the re-determining the preset brightness threshold of the fluorescence image comprises at least one of the following: re-determining the preset brightness threshold according to that part of the effective fluorescence signal does not correspond to the cell position identified in the phase difference image; and re-determining the preset brightness threshold according to that part of the non-effective fluorescence signal with brightness less than the preset brightness threshold in the fluorescence image corresponds to at least one part of the cell position in the phase difference image.

[0025] A third aspect of the present disclosure provides a transfection efficiency calculation device, comprising a processor, a memory for storing instructions executable by the processor, wherein when the instructions are executed by the processor, the processor is configured to perform the transfection efficiency calculation method of the second aspect of the present disclosure.

[0026] A fourth aspect of the present disclosure provides a computer readable storage medium having instructions stored thereon, when the instructions are executed by a processor, the processor is configured to perform the transfection efficiency calculation method of the second aspect of the present disclosure.

[0027] A fifth aspect of the present disclosure provides a microscope, comprising the transfection efficiency calculation device of the first aspect of the present disclosure.

[0028] The present disclosure provides a transfection efficiency calculation method and device, obtaining a phase difference image and a fluorescence image of a cell sample; identifying cells in the phase difference image and calculating the total number of cells; determining a preset brightness threshold of the fluorescence image, and filtering out effective fluorescence signals in the fluorescence image with brightness greater than or equal to the preset brightness threshold; comparing the phase difference image and the fluorescence image to identify transfection cells, which are cells corresponding to the position of the effective fluorescence signals; and calculating the transfection efficiency according to the total number of cells and the number of transfection cells. Thus, the present disclosure provides a technical solution that can accurately identify the total number of cells and the number of transfection cells, and accurately calculate the transfection efficiency, thereby solving the problem of low accuracy in identifying transfection cells and calculating transfection efficiency using a pure artificial intelligence solution in the related art.

[0029] Additional aspects and advantages of the present disclosure will be in part apparent and in part pointed out hereinafter. BRIEF DESCRIPTION OF DRAWINGS

[0030] The above and / or additional aspects and advantages of the present disclosure will become apparent and be readily appreciated from the following description, taken in conjunction with the accompanying drawings, in which:

[0031] FIG. 1 is a flowchart of a transfection efficiency calculation method according to an embodiment of the present disclosure.

[0032] FIG. 2 is a flowchart of another method for calculating transfection efficiency according to an embodiment of the present disclosure.

[0033] FIG. 3 is a flowchart of yet another method for calculating transfection efficiency according to an embodiment of the present disclosure.

[0034] FIG. 4 is a phase contrast image of a cell sample according to an embodiment of the present disclosure.

[0035] FIG. 5 is a fluorescence image of a cell sample according to an embodiment of the present disclosure.

[0036] FIG. 6 is a superimposed image according to an embodiment of the present disclosure.

[0037] FIG. 7 is a structural diagram of a device for calculating transfection efficiency according to an embodiment of the present disclosure.

[0038] FIG. 8 is a structural diagram of a system for implementing a method for calculating transfection efficiency according to an embodiment of the present disclosure. DETAILED DESCRIPTION

[0039] Embodiments of the present disclosure are described in detail below with reference to the accompanying drawings, in which like or similar elements are denoted by the same or similar reference numerals, and the embodiments described below are examples for explaining the present disclosure and should not be construed as limiting the present disclosure.

[0040] It should be noted that the terms "first", "second", and the like in the description and claims of the present disclosure and the above-described accompanying drawings are used to distinguish similar objects, and do not necessarily have to be used to describe a specific order or sequence. It should be understood that the data thus used can be interchanged under appropriate circumstances so that the embodiments of the present disclosure described herein can be implemented in an order other than that illustrated or described herein. The embodiments described in the following exemplary embodiments do not represent all embodiments consistent with the present disclosure. Rather, they are merely examples of apparatuses and methods consistent with some aspects of the present disclosure as detailed in the appended claims.

[0041] Cell transfection refers to a technique for introducing exogenous molecules (such as DNA, RNA, etc.) into eukaryotic cells. Transfection efficiency (transfection success rate) refers to the percentage of successfully transfected cells in the total number of cell samples. Therefore, to calculate the transfection efficiency (transfection success rate) of cells, two data are usually required: the total number of cells and the number of transfected cells.

[0042] Artificial intelligence solutions have relatively high accuracy in analyzing cell phase contrast images and accurately identifying the total number of cells. However, relying solely on artificial intelligence solutions cannot accurately identify the number of transfected cells, resulting in low accuracy of the calculated transfection efficiency and inconvenience to users.

[0043] To this end, the application provides a transfection efficiency calculation device, method, microscope and computer readable storage medium. According to a preset brightness threshold, effective fluorescent signals with brightness greater than or equal to the preset brightness threshold in a fluorescent image are filtered out. Then, by comparing the phase contrast image and the fluorescent image, transfection cells can be accurately identified. The transfection cells are cells corresponding to the positions of the effective fluorescent signals. Then, according to the total number of cells and the number of transfection cells, the transfection efficiency can be accurately calculated. Thus, the problem that transfection cells cannot be accurately identified by a simple artificial intelligence solution or a traditional image processing algorithm and the problem of low transfection efficiency calculation accuracy are effectively solved.

[0044] The transfection efficiency calculation method and device provided by the application will be described in detail below with reference to the drawings. It should be noted that the transfection efficiency calculation device provided by the application can be configured to execute the transfection efficiency calculation method provided by the application. The transfection efficiency calculation method will be described in detail below, and the description of the transfection efficiency calculation device can refer to the description of the transfection efficiency calculation method below.

[0045] FIG. 1 shows a flowchart of a transfection efficiency calculation method according to an embodiment of the present disclosure. The method can include but is not limited to the following steps:

[0046] In step S101, a phase contrast image and a fluorescent image of a cell sample are obtained.

[0047] The phase contrast image can be used to view cells that are not conspicuous and transparent in the bright field with high contrast and rich details. The living cells in the phase contrast image have clear edges and complete shapes. Therefore, by analyzing the phase contrast image, the living cells, i.e., effective cells, in the cell sample can be accurately identified.

[0048] The fluorescent image can display substances with fluorescent labels. The display result is the distribution of fluorescent signals in the fluorescent image.

[0049] In some embodiments, the phase contrast image and the fluorescent image of the cell sample are obtained by using different imaging modes of a microscope. For example, the microscope has a phase contrast imaging mode and a fluorescent imaging mode. In the phase contrast imaging mode, the microscope captures the cell sample to obtain the phase contrast image. In the fluorescent imaging mode, the microscope captures the cell sample to obtain the fluorescent image.

[0050] In other embodiments, the phase contrast image and the fluorescent image of the cell sample are obtained by using different lenses of a microscope or by using different microscopes. For example, the phase contrast image is obtained by using a phase contrast microscope, and the fluorescent image is obtained by using a fluorescent microscope.

[0051] It should be noted that in other embodiments, the phase contrast image and the fluorescent image of the cell sample can also be obtained by other techniques known in the art, and the present disclosure does not limit this.

[0052] In step S102, cells in the phase contrast image are identified and a total number of cells is calculated.

[0053] The total number of cells is the number of cells identified in the phase contrast image, wherein the identified cells are generally live cells.

[0054] In some embodiments, the identifying cells in the phase contrast image and calculating the total number of cells comprises determining a feature threshold of the cells, and using artificial intelligence deep learning capability to identify cells in the phase contrast image that meet a feature condition according to the feature threshold, the feature condition being that a corresponding feature value of the cell is greater than or equal to the feature threshold, or the feature condition being that the corresponding feature value of the cell is less than or equal to the feature threshold.

[0055] The feature value is at least one of a cell diameter, a cell radius, and a cell area. The artificial intelligence deep learning capability can more accurately identify cells in the phase contrast image and count the identified cells.

[0056] In some alternative embodiments, a preset maximum diameter / preset maximum radius of the cells is determined, and cells in the phase contrast image that have a maximum diameter less than or equal to the preset maximum diameter, or cells that have a maximum radius less than or equal to the preset maximum radius, are identified.

[0057] For example, the preset maximum diameter / preset maximum radius of the cells is determined according to user operation. The user operation includes, but is not limited to, inputting on an operation page, and determining the preset maximum diameter / preset maximum radius of the cells according to the input operation.

[0058] For example, the user inputs a preset maximum diameter of 70 pixels in a preset maximum diameter input box on the operation page, so that the preset maximum diameter of the cells expected by the user is determined to be 70 pixels.

[0059] For example, the user can manually measure the diameter of the cells in the phase contrast image using a ruler to help determine the preset maximum diameter.

[0060] In some alternative embodiments, a preset area of the cells is determined, and individual cells in the phase contrast image that have an area less than or equal to the preset area are identified.

[0061] In some alternative embodiments, the artificial intelligence deep learning capability can identify the cells in the phase contrast image according to deep learning experience.

[0062] The transfection efficiency calculation method provided by the embodiments of the present disclosure is executed by a transfection efficiency calculation device. The transfection efficiency calculation device can be communicatively connected with a display, and the display is used to display the phase contrast image.

[0063] In some embodiments, the identifying the cells in the phase contrast image and calculating the total number of cells comprises: identifying the cells in the phase contrast image, marking the outer contour of the cells with a closed contour line, and / or marking the cells with a cell layer.

[0064] For example, the cells in the phase contrast image are identified, and the outer contour of the cells is marked with a closed contour line. The user can observe the phase contrast image from the display, and the outer contour of the identified cells is marked with a contour line, so that the user can intuitively observe the outer contour of the identified cells, and then facilitate the user to judge the accuracy of cell identification.

[0065] For example, the cells in the phase contrast image are identified, and the cells are marked with a cell layer. The outer edge of the cell coating is substantially flush with the outer contour of the cell. The user can observe the phase contrast image from the display, and the identified cells are marked with a cell layer, so that the user can intuitively observe the identified cells, and then facilitate the user to judge the accuracy of cell identification.

[0066] For example, the cells in the phase contrast image are identified, and the outer contour of the cells is marked with a closed contour line, and the cells are marked with a cell layer. The user can observe the phase contrast image marked with the contour line and the cell coating from the display.

[0067] FIG. 4 shows a phase contrast image in which the identified cells are marked with a closed contour line and a corresponding cell layer according to an embodiment of the present disclosure, wherein the cell layer is filled in the closed contour line. The user can observe the marked phase contrast image and judge whether the cell identification is accurate. For example, if the user observes that the shape of a "cell" marked with a contour line and a cell coating is suspected to be a bubble, the cells in the phase contrast image can be re-identified by adjusting the identification parameters such as cell diameter or area to eliminate the possibility of identifying the bubble as a cell, improve the accuracy of the total number of cells calculation, and then improve the accuracy of the transfection efficiency calculation.

[0068] In step S103, a preset brightness threshold of the fluorescence image is determined, and the effective fluorescence signal with a brightness greater than or equal to the preset brightness threshold in the fluorescence image is filtered out.

[0069] Considering that the brightness of the fluorescence signal in the fluorescence image has differences, in the embodiments of the present disclosure, according to the preset brightness threshold, the fluorescence signal with a brightness greater than or equal to the preset brightness threshold in the fluorescence image is considered as an effective fluorescence signal, and the fluorescence signal with a brightness less than the preset brightness threshold is considered as a non-effective fluorescence signal. When identifying the transfected cells, only considering the effective fluorescence signal can improve the accuracy of the transfected cell identification.

[0070] In some embodiments, determining the preset luminance threshold of the fluorescent image comprises at least one of:

[0071] determining the preset luminance threshold according to a user operation;

[0072] determining the preset luminance threshold according to the fluorescent signals in the fluorescent image.

[0073] In some optional embodiments, the preset luminance threshold is determined according to a user operation, wherein the user operation comprises at least one of:

[0074] a sliding operation on a slider on a luminance bar;

[0075] an input operation.

[0076] For example, the user can input 66 in the preset luminance threshold input box in the operation page, so that the user-desired preset luminance threshold is determined to be 66. In the fluorescent image, the fluorescent signals with luminance greater than or equal to 66 are valid fluorescent signals, and the fluorescent signals with luminance less than 66 are invalid fluorescent signals, and only the valid fluorescent signals are filtered out.

[0077] For example, the user can slide the slider on the luminance bar in the operation page, and determine the user-desired preset luminance threshold by adjusting the position of the slider on the luminance bar. The adjustment range of the slider on the luminance bar is 0-255, for example, moving the slider from left to right, the preset luminance threshold increases from 0 to 255.

[0078] In some optional embodiments, determining the preset luminance threshold according to the fluorescent signals in the fluorescent image comprises: analyzing the distribution of the fluorescent signals in the fluorescent image based on an algorithm, and statistically determining a suitable preset luminance threshold according to the distribution of the fluorescent signals.

[0079] In some optional embodiments, determining the preset luminance threshold of the fluorescent image comprises:

[0080] statistically determining a first preset luminance threshold according to the fluorescent signals in the fluorescent image;

[0081] determining a second preset luminance threshold according to a user operation;

[0082] filtering out valid fluorescent signals in the fluorescent image with luminance greater than or equal to the second preset luminance threshold.

[0083] For example, the user judges that the first preset luminance threshold statistically determined by the algorithm is inaccurate according to experience, and inputs the desired second preset luminance threshold in the input box of the operation page, so that the second preset luminance threshold input by the user is the determined preset luminance threshold, and the valid fluorescent signals in the fluorescent image with luminance greater than or equal to the second preset luminance threshold are filtered out.

[0084] The transfection efficiency calculation method provided by the embodiments of the present disclosure is executed by a transfection efficiency calculation device. The transfection efficiency calculation device can be communicatively connected with a display, and the display is used to display the fluorescence image.

[0085] In some embodiments, the method further comprises determining a preset brightness threshold of the fluorescence image, and filtering out the effective fluorescent signal with a brightness greater than or equal to the preset brightness threshold in the fluorescence image.

[0086] The fluorescence region where the effective fluorescent signal is located is marked with a first color, and the first color is different from a second color of other fluorescence regions.

[0087] The fluorescence region where the effective fluorescent signal is located is an effective fluorescence region, and the other fluorescence regions are fluorescence regions where non-effective fluorescent signals are located, i.e., non-effective fluorescence regions.

[0088] The fluorescence region where the effective fluorescent signal is located is marked with a first color, and the color (second color) of the fluorescence region where the non-effective fluorescent signal is located is distinguished. The marked fluorescence image is displayed on the display, so that the user can observe the fluorescence region where the effective fluorescent signal is located.

[0089] For example, before the filtering, the fluorescence signal of the fluorescence image is displayed as green, and after the filtering, the fluorescence region where the effective fluorescent signal is located is displayed as purple, and there is a relatively obvious difference between the purple region and the other green regions. For example, FIG. 5 shows a fluorescence image of a cell sample according to an embodiment of the present disclosure, in which the fluorescence region where the effective fluorescent signal is located is marked as purple, and the other fluorescence regions are green.

[0090] When the preset brightness threshold is increased, the user can observe from the fluorescence image that the area of the fluorescence region marked with the first color gradually decreases. When the preset brightness threshold is decreased, the user can observe from the fluorescence image that the area of the fluorescence region marked with the first color gradually increases.

[0091] In some embodiments, the fluorescence regions marked with the first color have equal brightness and the brightness is greater than or equal to the preset brightness threshold.

[0092] The fluorescence region where the effective fluorescent signal is located is marked with the first color, and the brightness of the first color is set to be consistent, and the brightness of the first color is greater than or equal to the preset brightness threshold, so that the contrast between the fluorescence region where the effective fluorescent signal is located and the fluorescence region where the non-effective fluorescent signal is located can be increased, and the user can be facilitated to observe.

[0093] In step S104, the difference image and the fluorescence image are compared to identify the transfected cells, which are cells corresponding to the positions of the effective fluorescent signals.

[0094] The transfected cells are identified based on the cells identified in the phase contrast image and the effective fluorescent signals filtered out in the fluorescent image. The transfected cells are cells corresponding to the positions of the effective fluorescent signals in the phase contrast image.

[0095] It should be noted that, in order to effectively identify the transfected cells, the phase contrast image and the fluorescent image should be obtained by photographing the cell sample based on the same range size, that is, the positions of the cells in the phase contrast image and the positions of the cells in the fluorescent image should be the same, so that the transfected cells corresponding to the positions of the effective fluorescent signals can be accurately identified when the phase contrast image and the fluorescent image are compared.

[0096] In some embodiments, the transfected cells are cells corresponding to the positions of the fluorescent regions where the effective fluorescent signals are located, and the areas of the fluorescent regions corresponding to each transfected cell are greater than or equal to a preset area threshold. The fluorescent region where the effective fluorescent signal is located is an effective fluorescent region.

[0097] Specifically, the method comprises:

[0098] determining a preset area threshold;

[0099] comparing the phase contrast image and the fluorescent image to identify cells corresponding to the positions of the effective fluorescent regions and having areas of the corresponding effective fluorescent region positions greater than or equal to the preset area threshold. Such cells are transfected cells.

[0100] The preset area threshold can be preset.

[0101] For example, the preset area threshold is set to 2 pixel points, and when identifying the transfected cells, only the cells corresponding to the effective fluorescent regions with areas greater than or equal to 2 pixel points are identified as transfected cells. If a cell has a corresponding relationship with the position of an effective fluorescent region, but the total area of the corresponding effective fluorescent region is less than 2 pixel points, the cell cannot be identified as a transfected cell. For example, a dust area is identified as an effective fluorescent region, and when comparing the phase contrast image and the fluorescent image, it is found that there is a cell corresponding to the effective fluorescent region of the dust, but since the total area of the effective fluorescent region where the dust is located is less than 2 pixel points, the cell corresponding to the dust cannot be identified as a transfected cell.

[0102] It should be noted that the preset area threshold can be adjusted according to the identification accuracy of the transfected cells. For example, the preset area threshold can be 1 pixel point, or 3 pixel points, or other area values, and the present disclosure does not limit this.

[0103] In the embodiments of the present disclosure, the area of the effective fluorescence region corresponding to the transfected cells is greater than or equal to the preset area threshold, which effectively improves the recognition accuracy of the transfected cells.

[0104] In some embodiments, the phase difference image and the fluorescence image are superimposed to obtain a superimposed image.

[0105] The superimposed image can display image information of the phase difference image or image information of the fluorescence image. The transfected cells are the cells that display effective fluorescence signals in the superimposed image, that is, the cells that have superimposition with the effective fluorescence signals.

[0106] The transfected efficiency calculation method provided by the embodiments of the present disclosure is executed by a transfected efficiency calculation device. The transfected efficiency calculation device can be in communication connection with a display, and the display is used to display the superimposed image, so that the user can intuitively observe the comparison result of the phase difference image and the fluorescence image.

[0107] In some embodiments, the superimposed image is the result of algorithm superimposition of the phase difference image and the fluorescence image.

[0108] In some embodiments, the superimposed image displays or does not display the contour line and / or the cell layer.

[0109] For example, the superimposed image in FIG. 6 displays the contour line and the cell layer. In the superimposed image in FIG. 6, the effective fluorescence region marked as purple can also be seen, and the cells that have overlap with the purple region are identified as transfected cells.

[0110] For example, on the basis of FIG. 6, the user can make the superimposed image not display the contour line and the cell layer by operation, and then the superimposed effect of the original phase difference image and the fluorescence image of the cells can be observed, which helps the user to determine whether the determined preset brightness threshold is appropriate.

[0111] The superimposed image of the phase difference image and the fluorescence image is displayed to the user through the display, so that the user can intuitively observe the comparison result of the phase difference image and the fluorescence image, to determine whether the determined preset brightness threshold is appropriate, and to determine whether the recognition result of the transfected cells is accurate.

[0112] In some embodiments, if the user determines that the recognition result of the transfected cells is not accurate according to the superimposed image, the preset brightness threshold of the fluorescence image is re-determined, and the phase difference image and the fluorescence image are re-compared to identify the transfected cells. If the user determines that the recognition result of the transfected cells is accurate according to the superimposed image, the next step is continued.

[0113] Optionally, if the user observes from the superimposed image that part of the effective fluorescent signal does not correspond to the position of the cell identified in the phase difference image, or non-effective fluorescent signal with a luminance less than the preset luminance threshold corresponds to at least part of the position of the cell in the phase difference image, it is determined that the identification result of the transfected cell is inaccurate.

[0114] For example, the re-determination of the preset luminance threshold of the fluorescent image includes at least one of the following:

[0115] According to the part of the effective fluorescent signal does not correspond to the position of the cell identified in the phase difference image, the preset luminance threshold is re-determined;

[0116] According to the non-effective fluorescent signal with a luminance less than the preset luminance threshold corresponds to at least part of the position of the cell in the phase difference image, the preset luminance threshold is re-determined.

[0117] Optionally, if the user observes from the superimposed image that part of the effective fluorescent region is located outside the identified cell region, it is possible that the preset luminance threshold is set too small, and the effective fluorescent region is not reasonably delimited, and the preset luminance threshold can be appropriately increased.

[0118] Optionally, if the user observes from the superimposed image that the non-effective fluorescent region is superimposed on the position of the identified cell, and this part of the cell is not identified as a transfected cell, it is possible that the preset luminance threshold is set too large, and the effective fluorescent region is not reasonably delimited, and the preset luminance threshold can be appropriately reduced.

[0119] The user can adjust the preset luminance threshold while observing the superimposed image to determine a suitable preset luminance threshold. For example, the slider on the luminance bar can be adjusted while observing the superimposed image to observe the change in the correspondence between the effective fluorescent region and the cell to determine a suitable preset luminance threshold.

[0120] For example, as shown in FIG. 6, the effective fluorescent region (purple fluorescent region) is basically located within the cell region, and the identified cell basically does not have a corresponding relationship with the non-effective fluorescent region (green fluorescent region), and therefore it can be considered that in the embodiment shown in FIG. 6, the preset luminance threshold is accurately set, and the calculated transfection efficiency is also accurate.

[0121] In step S105, the total number of cells is the total number of cells in step S102, and the number of transfected cells is the number of transfected cells identified in S104, and the transfection efficiency is calculated according to the total number of cells and the number of transfected cells.

[0122] In step S105, the total number of cells is the total number of cells in step S102, and the number of transfected cells is the number of transfected cells identified in S104, and the transfection efficiency is calculated according to the total number of cells and the number of transfected cells.

[0123] In some embodiments, as shown in FIG. 6, the transfection efficiency is displayed on a display page of the display.

[0124] According to the transfection efficiency calculation method provided by the embodiment of the present disclosure, the phase contrast image and the fluorescence image of the cell sample are obtained; the cells in the phase contrast image are identified and the total number of cells is calculated; the preset brightness threshold of the fluorescence image is determined, and the effective fluorescent signal with brightness greater than or equal to the preset brightness threshold in the fluorescence image is filtered out; the phase contrast image and the fluorescence image are compared to identify the transfected cells, which are the cells corresponding to the position of the effective fluorescent signal; and the transfection efficiency is calculated according to the total number of cells and the number of transfected cells. Thus, the present disclosure provides a technical solution that can accurately identify the total number of cells and the number of transfected cells, and then accurately calculate the transfection efficiency, solving the problem of low accuracy of transfection cell identification and transfection efficiency calculation in the related art using a pure artificial intelligence solution.

[0125] The information processing method related to the embodiment of the present disclosure can include at least one of steps S101-S105.

[0126] In the embodiment of the present disclosure, part or all of the steps, and the optional implementation manners thereof, can be combined with part or all of the steps in other embodiments, or can be combined with the optional implementation manners of other embodiments.

[0127] FIG. 2 shows a flowchart of another transfection efficiency calculation method according to an embodiment of the present disclosure. The transfection efficiency calculation method provided by the embodiment of the present disclosure is executed by a transfection efficiency calculation device. The transfection efficiency calculation device can be in communication connection with a display, and the display is used to display at least one of the phase contrast image, the fluorescence image, the superimposed image, and the transfection efficiency. As shown in FIG. 2, the method can include but is not limited to the following steps:

[0128] Step S201, obtaining a phase contrast image and a fluorescence image of a cell sample.

[0129] For detailed introduction of step S201, reference can be made to the description of step S101 in the above-mentioned embodiments, which will not be repeated here.

[0130] Step S202, identifying the cells in the phase contrast image and calculating the total number of cells, using a closed contour line to mark the outer contour of the cells, and using a cell layer to mark the cells.

[0131] In some embodiments, the display displays the phase contrast image, and the phase contrast image displays or does not display the contour line and the cell layer. For example, as shown in FIG. 4, the phase contrast image displays the contour line and the cell layer, and the user observes the phase contrast image marked with the contour line and the cell layer from the display to determine whether the cell identification is accurate.

[0132] For details of step S202, refer to the description of step S102 in the above embodiments, which will not be repeated here.

[0133] At step S203, a preset luminance threshold of the fluorescent image is determined, and effective fluorescent signals in the fluorescent image with luminance greater than or equal to the preset luminance threshold are filtered out.

[0134] In some embodiments, the display displays the fluorescent image, as shown in FIG. 5, in which the fluorescent region corresponding to the effective fluorescent signals is marked in a first color, and the fluorescent region corresponding to the non-effective fluorescent signals is displayed in a second color, and the first color is different from the second color.

[0135] For details of step S203, refer to the description of step S103 in the above embodiments, which will not be repeated here.

[0136] At step S204, the phase contrast image and the fluorescent image are superimposed to obtain a superimposed image.

[0137] In some embodiments, the display displays the superimposed image, and the superimposed image displays or does not display the contour line and the cell layer. For example, as shown in FIG. 6, the superimposed image displays the contour line and the cell layer.

[0138] By displaying the superimposed image of the phase contrast image and the fluorescent image to the user, the user can intuitively observe the comparison result of the phase contrast image and the fluorescent image to determine whether the preset luminance threshold determined in step S203 is appropriate.

[0139] For example, if the user determines that the transfection cell recognition result is inaccurate according to the superimposed image, the preset luminance threshold of the fluorescent image is re-determined.

[0140] For example, re-determining the preset luminance threshold of the fluorescent image includes at least one of the following:

[0141] According to the fact that part of the effective fluorescent signals do not correspond to the positions of the cells recognized in the phase contrast image, the preset luminance threshold is re-determined;

[0142] According to the fact that the non-effective fluorescent signals with luminance less than the preset luminance threshold in the fluorescent image correspond to at least part of the positions of the cells in the phase contrast image, the preset luminance threshold is re-determined.

[0143] The user can adjust the preset luminance threshold while observing the superimposed image to determine an appropriate preset luminance threshold. For example, the slider on the luminance bar can be adjusted while observing the superimposed image to observe the change in the correspondence between the effective fluorescent region and the cells, so as to determine an appropriate preset luminance threshold.

[0144] Step S205, comparing the phase contrast image and the fluorescence image to identify the transfected cells, which are the cells corresponding to the positions of the effective fluorescence signals.

[0145] In some embodiments, the transfected cells are identified by comparing the phase contrast image and the fluorescence image through an algorithm.

[0146] It should be noted that the transfected cells can also be identified based on the superimposed image in step S204 by comparing the phase contrast image and the fluorescence image.

[0147] In general, the transfected cells are identified by comparing the phase contrast image and the fluorescence image through an algorithm.

[0148] For detailed description of step S205, reference can be made to the description of step S104 in the above embodiments, which will not be repeated here.

[0149] Step S206, calculating the transfection efficiency according to the total number of cells and the number of transfected cells.

[0150] In some embodiments, the display displays the transfection efficiency.

[0151] For detailed description of step S206, reference can be made to the description of step S105 in the above embodiments, which will not be repeated here.

[0152] According to the transfection efficiency calculation method provided by the embodiments of the present disclosure, the total number of cells and the number of transfected cells can be accurately identified, and the transfection efficiency can be accurately calculated, thereby solving the problem that the pure artificial intelligence solution calculation method in the related art cannot accurately identify the transfected cells and the transfection efficiency calculation accuracy is low.

[0153] FIG. 3 shows a flowchart of another transfection efficiency calculation method according to an embodiment of the present disclosure. As shown in FIG. 3, the method can include but is not limited to the following steps:

[0154] Step S301, obtaining a phase contrast image and a fluorescence image of a cell sample.

[0155] For detailed description of step S301, reference can be made to the description of step S101 in the above embodiments, which will not be repeated here.

[0156] Step S302, identifying the cells in the phase contrast image and calculating the total number of cells.

[0157] For detailed description of step S302, reference can be made to the description of step S102 in the above embodiments, which will not be repeated here.

[0158] Step S303, determining a preset brightness threshold of the fluorescence image, and filtering out the effective fluorescence signals in the fluorescence image with brightness greater than or equal to the preset brightness threshold.

[0159] The detailed description of step S303 can refer to the description of step S103 in the above embodiment, and will not be repeated here.

[0160] Step S304, comparing the phase contrast image and the fluorescence image to identify the transfected cells, which are the cells corresponding to the positions of the effective fluorescence signals.

[0161] The detailed description of step S304 can refer to the description of step S104 in the above embodiment, and will not be repeated here.

[0162] Step S305, determining whether the transfected cells identified in step S304 are accurate.

[0163] If it is determined that the identified transfected cells are not accurate, then:

[0164] re-determining the preset luminance threshold of the fluorescence image; and

[0165] re-comparing the phase contrast image and the fluorescence image to identify the transfected cells;

[0166] If it is determined that the identified transfected cells are accurate, then step S306 is performed.

[0167] That is, when it is determined that the transfected cells identified in step S304 are not accurate, steps S303 and S304 are re-executed.

[0168] In some embodiments, artificial intelligence is used to determine whether the transfected cells identified in step S304 are accurate.

[0169] For example, if the artificial intelligence identifies that part of the effective fluorescence signals do not correspond to the positions of the cells identified in the phase contrast image, or identifies that the non-effective fluorescence signals with luminance less than the preset luminance threshold in the fluorescence image correspond to at least part of the positions of the cells in the phase contrast image, it is determined that the identified transfected cells are not accurate.

[0170] For example, the preset luminance threshold is re-determined according to that part of the effective fluorescence signals do not correspond to the positions of the cells identified in the phase contrast image, or the preset luminance threshold is re-determined according to that the non-effective fluorescence signals with luminance less than the preset luminance threshold in the fluorescence image correspond to at least part of the positions of the cells in the phase contrast image.

[0171] Step S306, calculating the transfection efficiency according to the total number of cells and the number of transfected cells.

[0172] The detailed description of step S306 can refer to the description of step S105 in the above embodiment, and will not be repeated here.

[0173] According to the transfection efficiency calculation method provided by the embodiment of the present disclosure, the total number of cells and the number of transfected cells can be accurately identified, and the transfection efficiency can be accurately calculated, thereby solving the problem that the pure artificial intelligence solution calculation method in the related art cannot accurately identify the transfected cells and the transfection efficiency calculation accuracy is low.

[0174] The embodiment of the present disclosure also proposes a device for implementing any of the above methods, for example, a device including units or modules for implementing each step in any of the above methods.

[0175] It should be understood that the division of each unit or module in the device is only a logical function division, and all or part of the units or modules can be integrated into one physical entity or physically separated in actual implementation. In addition, the units or modules in the device can be implemented in the form of processor calling software: for example, the device includes a processor connected with a memory, the memory stores instructions, and the processor calls the instructions stored in the memory to implement any of the above methods or to implement the functions of each unit or module of the device, wherein the processor is, for example, a general processor such as a central processing unit (CPU) or a microprocessor, and the memory is a memory in the device or a memory outside the device. Alternatively, the units or modules in the device can be implemented in the form of hardware circuit, and the functions of part or all of the units or modules can be implemented by designing the hardware circuit, and the hardware circuit can be understood as one or more processors; for example, in one implementation, the hardware circuit is an application-specific integrated circuit (ASIC), and the functions of part or all of the units or modules are implemented by designing the logical relationship of elements in the circuit; for another example, in another implementation, the hardware circuit is a programmable logic device (PLD), and taking a field programmable gate array (FPGA) as an example, it can include a large number of logic gate circuits, and the connection relationship between the logic gate circuits is configured by a configuration file, thereby implementing the functions of part or all of the units or modules. All units or modules of the above device can be implemented in the form of processor calling software, or all units or modules can be implemented in the form of hardware circuit, or part of the units or modules are implemented in the form of processor calling software, and the remaining part is implemented in the form of hardware circuit.

[0176] In the embodiments of the present disclosure, the processor is a circuit with signal processing capability. In one implementation, the processor can be a circuit with instruction reading and running capability, such as a central processing unit (CPU), a microprocessor, a graphics processing unit (GPU) (which can be understood as a microprocessor), a digital signal processor (DSP), or the like. In another implementation, the processor can implement certain functions through a logical relationship of hardware circuits, and the logical relationship of the hardware circuits is fixed or can be reconfigured. For example, the processor is a hardware circuit implemented by an application-specific integrated circuit (ASIC) or a programmable logic device (PLD), such as an FPGA. In the reconfigurable hardware circuit, the processor loads a configuration document to implement the configuration of the hardware circuit. It can be understood that the processor loads instructions to implement the functions of the above part or all units or modules. In addition, the hardware circuit can also be designed for artificial intelligence, which can be understood as an ASIC, such as a neural network processing unit (NPU), a tensor processing unit (TPU), a deep learning processing unit (DPU), or the like.

[0177] FIG. 7 is a structural schematic diagram of a transfection efficiency calculation device provided by an embodiment of the present disclosure. As shown in FIG. 7, the transfection efficiency calculation device 700 can include an acquisition module 701, a first identification module 702, a processing module 703, a second identification module 704, and a calculation module 705.

[0178] The acquisition module 701 is configured to acquire a phase contrast image and a fluorescence image of a cell sample.

[0179] The first identification module 702 is configured to identify cells in the phase contrast image and calculate a total number of the cells.

[0180] The processing module 703 is configured to determine a preset brightness threshold of the fluorescence image, and filter out effective fluorescence signals in the fluorescence image with a brightness greater than or equal to the preset brightness threshold.

[0181] The second identification module 704 is configured to compare the phase contrast image and the fluorescence image to identify transfection cells, the transfection cells being cells corresponding to the effective fluorescence signals.

[0182] The computing module 705 calculates the transfection efficiency according to the total number of cells and the number of transfected cells.

[0183] Optionally, the transfection efficiency calculation device 700 further comprises a superimposition module 706 configured to superimpose the phase contrast image and the fluorescence image to obtain a superimposed image.

[0184] Optionally, the transfected cells are cells corresponding to positions of the fluorescence regions where the effective fluorescence signals are located, and an area of each fluorescence region corresponding to a transfected cell is greater than or equal to a preset area threshold.

[0185] Optionally, the first identification module 702 is configured to identify the cells in the phase contrast image, identify the outer contour of the cells by using a closed contour line, and / or identify the cells by using a cell layer.

[0186] Optionally, the superimposed image displays or does not display the contour line and / or the cell layer.

[0187] Optionally, the first identification module 702 is configured to determine a feature threshold of the cells, and identify the cells in the phase contrast image that meet a feature condition by using an artificial intelligence deep learning capability according to the feature threshold, the feature condition being that a corresponding feature value of the cells is greater than or equal to the feature threshold or the corresponding feature value of the cells is less than or equal to the feature threshold, wherein the feature value is at least one of a cell diameter, a cell radius, and a cell area.

[0188] Optionally, the processing module 703 is configured to determine the preset brightness threshold according to a user operation, and / or determine the preset brightness threshold according to the fluorescence signals in the fluorescence image.

[0189] Optionally, the processing module 703 is configured to mark the fluorescence regions where the effective fluorescence signals are located by using a first color, the first color being different from a second color of other fluorescence regions.

[0190] Optionally, the fluorescence regions marked by using the first color have equal brightness and the brightness is greater than or equal to the preset brightness threshold.

[0191] Optionally, the transfection efficiency calculation device 700 further comprises a judgment module 707 configured to judge whether the identified transfected cells are accurate, if it is determined that the identified transfected cells are not accurate, the processing module 703 re-determines the preset brightness threshold of the fluorescence image, and the second identification module 704 re-compares the phase contrast image and the fluorescence image to identify the transfected cells, and if it is determined that the identified transfected cells are accurate, the computing module 705 calculates the transfection efficiency according to the total number of cells and the number of transfected cells.

[0192] Optionally, the processing module 703 is configured to: determine the preset luminance threshold again according to that the partial effective fluorescence signal does not correspond to the cell position identified in the phase contrast image; and / or determine the preset luminance threshold again according to that the non-effective fluorescence signal with luminance less than the preset luminance threshold in the fluorescence image corresponds to at least part of the cell position in the phase contrast image.

[0193] Optionally, the transfection efficiency calculation apparatus 700 further includes a display module configured to display at least one of the phase contrast image, the fluorescence image, the superimposed image and the transfection efficiency.

[0194] Optionally, the display module is a display.

[0195] The transfection efficiency calculation apparatus provided by the embodiments of the present disclosure can accurately identify the total number of cells and the number of transfected cells, and further accurately calculate the transfection efficiency, thereby solving the problem of low accuracy of the transfection efficiency calculation in the related art by using a pure artificial intelligence solution calculation method.

[0196] Some embodiments relate to a microscope. FIG. 8 shows a schematic diagram of a system 900 configured to perform the methods described herein. The system 900 comprises a microscope 910, a camera 920 and a computer system 930. The camera is configured to capture non-fluorescent images, the microscope 910 is configured to acquire fluorescent images, and is connected to the computer system 930. The computer system 930 is configured to perform at least part of the methods described herein. The computer system 930 can be configured to perform a machine learning algorithm. The computer system 930 and the microscope 910, the camera 920 can be separate entities, but can also be integrated into one common housing. The computer system 930 can be part of a central processing system of the microscope 910 and / or the computer system 930 and the camera 920 can be part of a subassembly of the microscope 910, such as a sensor, an actuator, a camera or an illumination unit of the microscope 910, etc.

[0197] The computer system 930 can be a local computer device (e.g., a personal computer, a notebook, a tablet, or a mobile phone) having one or more processors and one or more storage devices, or can also be a distributed computer system (e.g., having one or more processors and one or more storage devices distributed at various locations, such as a local client and / or one or more remote server sites and / or data centers). The computer system 930 can include any circuit or combination of circuits. In one embodiment, the computer system 930 can include one or more processors, which can be of any type. As used herein, a processor can include any type of computational circuit such as, for example, but not limited to, a microprocessor, a microcontroller, a complex instruction set computing (CISC) microprocessor, a reduced instruction set computing (RISC) microprocessor, a very long instruction word (VLIW) microprocessor, a graphics processor, a digital signal processor (DSP), a multi-core processor, a field programmable gate array (FPGA) such as a microscope or microscope assembly (e.g., a camera), or any other type of processor or processing circuit. Other types of circuit that can be included in the computer system 930 can be custom circuit, an application-specific integrated circuit (ASIC), etc., such as one or more circuits (e.g., a communication circuit) used in a wireless device such as a mobile phone, a tablet, a notebook, a two-way radio, and similar electronic systems. The computer system 930 can include one or more storage devices, which can include one or more storage elements suitable for a particular application, such as a main memory in the form of random access memory (RAM), one or more hard drives, and / or one or more drives that handle removable media such as compact disks (CDs), flash memory cards, digital video disks (DVDs), etc. The computer system 930 can also include a display device, one or more speakers, and a keyboard and / or controller, which can include a mouse, a trackball, a touch screen, a voice recognition device, or any other device that allows a system user to input information to and receive information from the computer system 930.

[0198] Some or all of the method steps can be performed by (or using) a hardware device (e.g., a processor, a microprocessor, a programmable computer or an electronic circuit). In some embodiments, such a device can perform one or more of the most important method steps.

[0199] Depending on certain implementation requirements, embodiments of the application can be implemented in hardware or in software. The implementation can be carried out using a non-transitory storage medium such as a digital storage medium, for example a floppy diskette, a DVD, a Blu-Ray, a CD, a ROM, a PROM, and EPROM, an EEPROM or a FLASH memory, having electronically readable control signals stored thereon, which cooperate (or are capable of cooperating) with a programmable computer system such that the respective method is performed. Therefore, the digital storage medium can be computer readable.

[0200] Some embodiments of the application comprise a data carrier having electronically readable control signals, which are capable of cooperating with a programmable computer system, such that one of the methods described herein can be performed.

[0201] Generally, embodiments of the present application can be implemented as a computer program product with a program code, the program code being operative for performing one of the methods when the computer program product runs on a computer. The program code can for example be stored on a machine readable carrier.

[0202] Other embodiments comprise the computer program for performing one of the methods described herein, stored on a machine readable carrier.

[0203] In other words, an embodiment of the inventive method is, therefore, a computer program having a program code for performing one of the methods described herein, when the computer program runs on a computer.

[0204] A further embodiment of the inventive methods is, therefore, a computer having installed thereon the computer program for performing one of the methods described herein.

[0205] A further embodiment of the inventive method is, therefore, a data stream or a sequence of signals representing the computer program for performing one of the methods described herein. The data stream or the sequence of signals can for example be configured to be transferred via a data communication connection, for example via the Internet.

[0206] A further embodiment comprises a processing means, for example a computer, or a programmable logic device, configured to or adapted for performing one of the methods described herein.

[0207] A further embodiment comprises a computer having installed thereon the computer program for performing one of the methods described herein.

[0208] Yet another embodiment of the present application includes an apparatus or system configured to transfer a computer program for performing one of the methods described herein (e.g., electronically or optically) to a receiver. The receiver could, for example, be a computer, mobile device, storage device, etc. The apparatus or system might include a file server for transferring the computer program to the receiver.

[0209] In some embodiments, a programmable logic device (e.g., a field programmable gate array) can be used to perform some or all of the functionalities of the methods described herein. In some embodiments, a field programmable gate array can cooperate with a microprocessor in order to perform one of the methods described herein. Generally, the methods are preferably performed by any hardware apparatus.

[0210] As used herein, the term "and / or" includes any and all combinations of one or more of the associated listed items and can be abbreviated as " / ".

[0211] Although some aspects have been described in the context of an apparatus, it is clear that other aspects also represent a description, although in an opposite direction, of a corresponding method, wherein a block or device corresponds to a method step or a feature of a method step. Analogously, aspects described in the context of a method step also represent a description of a corresponding block or item or feature of a corresponding apparatus, although in an opposite direction.

Claims

1. A device for calculating transfection efficiency, the device being configured to: obtain a phase contrast image and a fluorescence image of a cell sample; identify cells in the phase contrast image and calculate a total number of cells; determine a preset luminance threshold of the fluorescence image, and filter out effective fluorescent signals in the fluorescence image with luminance greater than or equal to the preset luminance threshold; compare the phase contrast image and the fluorescence image to identify transfected cells, the transfected cells being cells corresponding to positions of the effective fluorescent signals; and calculate the transfection efficiency according to the total number of cells and a number of the transfected cells. The transfected cells are cells corresponding to positions of fluorescent regions where the effective fluorescent signals are located, and an area of a fluorescent region corresponding to each of the transfected cells is greater than or equal to a preset area threshold. The device is configured to: identify cells in the phase contrast image, mark an outer contour of each of the cells using a closed contour line, and / or mark the cells using a cell layer. The device is configured to: superimpose the phase contrast image and the fluorescence image to obtain a superimposed image, the superimposed image including the contour line and / or the cell layer. The device is configured to: determine a feature threshold of the cells, and identify cells in the phase contrast image that satisfy a feature condition using artificial intelligence deep learning capability according to the feature threshold, the feature condition being that a corresponding feature value of the cells is greater than or equal to the feature threshold or less than or equal to the feature threshold, the feature value being at least one of a cell diameter, a cell radius and a cell area. The device is configured to: determine the preset luminance threshold according to a user operation; and / or determine the preset luminance threshold according to fluorescent signals in the fluorescence image. The device is configured to: mark the fluorescent regions where the effective fluorescent signals are located using a first color, the first color being different from a second color of other fluorescent regions.

2. The apparatus of claim 1, wherein, The device is configured to: determine whether the identified transfected cells are accurate: if it is determined that the identified transfected cells are not accurate, then: redetermine the preset luminance threshold of the fluorescence image; and re-compare the phase contrast image and the fluorescence image to identify transfected cells; and if it is determined that the identified transfected cells are accurate, then: calculate the transfection efficiency according to the total number of cells and the number of the transfected cells.

3. The apparatus of claim 1 or 2, wherein, The device is configured to perform at least one of the following: redetermine the preset luminance threshold according to a part of the effective fluorescent signals not corresponding to positions of the cells identified in the phase contrast image; and redetermine the preset luminance threshold according to non-effective fluorescent signals in the fluorescence image with luminance less than the preset luminance threshold corresponding to positions of at least a part of the cells in the phase contrast image. The method comprises: obtaining a phase contrast image and a fluorescence image of a cell sample; identifying cells in the phase contrast image and calculating a total number of cells; 4. The apparatus of claim 3, wherein, ​ ​ 5. The apparatus of any one of claims 1-4, wherein, ​ ​ 6. The apparatus of any one of claims 1-5, wherein, ​ ​ ​ 7. The apparatus of any one of claims 1-6, wherein, ​ ​ 8. The apparatus of claim 7, wherein, ​ 9. The apparatus of any one of claims 1-8, wherein, ​ ​ ​ ​ ​ ​ ​ 10. The apparatus of claim 9, wherein, ​ ​ ​ 11. A method of calculating transfection efficiency, wherein, ​ ​ ​ determining a preset brightness threshold of the fluorescent image, and filtering out effective fluorescent signals in the fluorescent image with brightness greater than or equal to the preset brightness threshold; comparing the phase contrast image and the fluorescent image to identify transfected cells, the transfected cells being cells corresponding to positions of the effective fluorescent signals; and calculating the transfection efficiency according to the total number of cells and the number of the transfected cells.

12. The method according to claim 11, wherein, The identifying the cells in the phase contrast image and calculating the total number of cells comprises: determining a feature threshold of the cells, and identifying cells in the phase contrast image that meet a feature condition according to the feature threshold and artificial intelligence deep learning capability, the feature condition being that a corresponding feature value of the cells is greater than or equal to the feature threshold or the corresponding feature value of the cells is less than or equal to the feature threshold, the feature value being at least one of a cell diameter, a cell radius and a cell area; The determining the preset brightness threshold of the fluorescent image comprises: determining the preset brightness threshold according to a user operation; and / or, determining the preset brightness threshold according to fluorescent signals in the fluorescent image; The determining the preset brightness threshold of the fluorescent image and filtering out the effective fluorescent signals in the fluorescent image with brightness greater than or equal to the preset brightness threshold further comprises: marking a fluorescent region where the effective fluorescent signals are located with a first color, the first color being different from a second color of other fluorescent regions.

13. The method according to claim 11 or 12, wherein, The identifying the transfected cells further comprises: judging whether the identified transfected cells are accurate: if it is determined that the identified transfected cells are not accurate, then: redetermining the preset brightness threshold of the fluorescent image; recomparing the phase contrast image and the fluorescent image to identify transfected cells; if it is determined that the identified transfected cells are accurate, then: calculating the transfection efficiency according to the total number of cells and the number of the transfected cells.

14. A microscope, characterized by comprises: The transfection efficiency calculation device according to any one of claims 1-10.

15. A computer readable storage medium having instructions stored thereon, when the instructions are executed by a processor, the processor is configured to perform the transfection efficiency calculation method according to any one of claims 11-13.

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