Information processing method and system for a flow cytometer and flow cytometer

Abstract

Disclosed are an information processing method and system for a flow cytometer and the flow cytometer. The information processing method comprises: generating an initial spillover matrix based on detection results of a plurality of single-stained samples input to the flow cytometer, wherein each single-stained sample is stained with a corresponding one of a plurality of fluorescent dyes; for each of the plurality of single-stained samples, de-mixing the detection results for the single-stained sample using the generated initial spillover matrix, generating a first scatter plot of a primary channel relative to each of a plurality of secondary channels based on the de-mixed results, and calculating a first slope of a straight line connecting a center of a positive cell population and a center of a negative cell population in each first scatter plot as a compensation gap; and providing a final spillover matrix for detecting an actual sample based on a plurality of compensation gaps calculated for the plurality of single-stained samples.

Description

TECHNICAL FIELD

[0001] The present disclosure relates to flow cytometry, and more particularly, to an information processing method and an information processing system for a flow cytometer, and a flow cytometer. BACKGROUND

[0002] Flow cytometry is a biological technique for counting and sorting micro-particles suspended in a fluid. In a typical flow cytometer, cells dyed with fluorescent dyes are irradiated with a proper light source, the cells emit fluorescence when excited, the fluorescence is collected and converted into an electrical signal by a photoelectric converter, and the electrical signal is input to a computer analyzer after amplification for quantitative analysis and sorting of the cells.

[0003] The cells to be detected are usually dyed with multiple fluorescent dyes, and different cells can be dyed with different fluorescent dyes due to the difference in the characteristics of the cells, so that different fluorescence is emitted when excited. By collecting and analyzing the overall fluorescence emitted by all cells when excited, the cells dyed with each fluorescent dye and the number thereof can be identified, thereby achieving sorting and counting of the cells. However, the overall fluorescence emitted by all cells contains multiple fluorescent dyes with different wavelengths, and the spectra of these fluorescent dyes are mixed together. In order to analyze, it is necessary to distinguish each fluorescent dye from each other, which is called unmixing. Spillover matrix can be used for unmixing.

[0004] In practice, before applying the generated spillover matrix to the actual cell sample to be detected, it is usually applied to a test sample to predict the quality of the result of unmixing using the spillover matrix. If the result of unmixing meets the requirements, the spillover matrix can be applied to the actual cell sample to be detected. Otherwise, it may be necessary to regenerate the spillover matrix.

[0005] In the conventional technology, the user often determines whether the result of unmixing meets the requirements by experience, so this method is subjective and may not be accurate enough due to the user's neglect of some details.

[0006] Although some indicators for helping the user to determine whether the result of unmixing meets the requirements have been proposed in recent years, such as Spillover Spreading Matrix (SSM), complexity indicator, similarity indicator, etc., these indicators only reflect the spreading caused by dye color or instrument noise and the similarity between dye colors, and cannot directly reflect the pros and cons of the result of unmixing. SUMMARY

[0007] The present disclosure aims to provide an information processing technique for a flow cytometer, which substantially avoids one or more problems caused by the limitations and shortcomings of the prior art.

[0008] According to one aspect of this disclosure, an information processing method for a flow cytometer is provided, comprising: generating an initial overflow matrix based on detection results of multiple single-stained samples input to the flow cytometer, wherein each single-stained sample is stained with a corresponding one of a plurality of fluorescent dyes; for each of the plurality of single-stained samples, unmixing the detection results for that single-stained sample using the generated initial overflow matrix; generating a first scatter plot of a main channel relative to each of a plurality of secondary channels based on the unmixing results; and calculating a first slope of a straight line connecting the center of a positive cell population and the center of a negative cell population in each of the first scatter plots as a compensation gap; and providing a final overflow matrix for detecting actual samples based on a plurality of compensation gaps calculated for the plurality of single-stained samples.

[0009] According to another aspect of this disclosure, an information processing system for a flow cytometer is provided, comprising a processing unit configured to: generate an initial overflow matrix based on detection results of a plurality of monostained samples input to the flow cytometer, wherein each monostained sample is stained with a corresponding one of a plurality of fluorescent dyes; for each of the plurality of monostained samples, unmix the detection results for that monostained sample using the generated initial overflow matrix; generate a first scatter plot of the main channel relative to each of the plurality of secondary channels based on the unmixing results; and calculate a first slope of the straight line connecting the center of the positive cell population and the center of the negative cell population in each of the first scatter plots as a compensation gap; and provide a final overflow matrix for detecting actual samples based on the plurality of compensation gaps calculated for the plurality of monostained samples.

[0010] According to another aspect of this disclosure, a flow cytometer including the aforementioned information processing system is provided.

[0011] According to another aspect of this disclosure, a non-transitory computer-readable medium storing instructions that, when executed by a processor, cause the processor to perform the aforementioned information processing method.

[0012] In accordance with other aspects of this disclosure, computer program code and computer program products for implementing the above information processing methods are also provided. Attached Figure Description

[0013] Figure 1 A flowchart of the overall method for generating an overflow matrix according to this disclosure is shown.

[0014] Figure 2 A flowchart illustrating a method for generating an initial overflow matrix according to this disclosure is shown.

[0015] Figure 3The selection of a target cell population in an FSC-SSC scatter plot is illustrated schematically.

[0016] Figure 4 The diagram illustrates the selection of negative and positive cells in a histogram.

[0017] Figure 5 A scatter plot of the main channel relative to one of the secondary channels is shown schematically.

[0018] Figure 6 A flowchart illustrating a method for obtaining and analyzing unmixed results according to this disclosure is shown.

[0019] Figure 7 A scatter plot of the main channel relative to the two secondary channels is shown schematically.

[0020] Figure 8 A user interface for generating an initial overflow matrix, according to this disclosure, is illustrated schematically.

[0021] Figure 9 The user interface for viewing the unmixing results according to this disclosure is illustrated schematically.

[0022] Figure 10A and Figure 10B The user interface for viewing a scatter plot according to this disclosure is illustrated schematically.

[0023] Figures 11A-11C A user interface for setting the gain value of a channel according to this disclosure is illustrated schematically.

[0024] Figure 12 A user interface for setting channels for the coordinate axes of a graph according to this disclosure is illustrated schematically.

[0025] Figure 13 The user interface for setting up channels on an oscilloscope according to this disclosure is illustrated schematically.

[0026] Figure 14 An exemplary configuration block diagram of computer hardware implementing the present disclosure is shown. Detailed Implementation

[0027] The specific implementation methods according to this disclosure are described in detail below with reference to the accompanying drawings.

[0028] Figure 1 A flowchart illustrating the overall method for generating an overflow matrix according to this disclosure is shown. Figure 1 As shown, in step S110, a single-stained sample is input into the flow cytometer, and an initial overflow matrix is ​​generated based on the detection results of the fluorescence emitted by the single-stained sample by the flow cytometer.

[0029] In step S120, the flow cytometer inputs the single-stained sample used in step S110, and unmixes the detection results for that single-stained sample using the generated initial spillover matrix. Then, in step S130, the obtained unmixing results are analyzed. Based on the analysis results, in step S140, a final spillover matrix for detecting actual cell samples is provided.

[0030] The following will combine Figures 2-5 Step S110 is described in detail.

[0031] Figure 2 A flowchart illustrating a method for generating an initial overflow matrix according to this disclosure is shown. Figure 2 As shown, in step S210, a single-stained cell sample stained with the j-th fluorescent dye is input into a flow cytometer, where j = 1, 2, ..., P. Since the various cells in the sample have different characteristics, some cells in the single-stained sample are stained with the fluorescence corresponding to the j-th dye and are excited by light in the flow cytometer to emit the corresponding fluorescence, while other cells are not stained with the j-th dye and therefore are not excited to emit the corresponding fluorescence in the flow cytometer.

[0032] After processing by flow cytometry, FSC-SSC scatter plots of single-stained samples can be generated. FSC-SSC scatter plots can be used to group cells based on cell size and granularity, such as lymphocytes, neutrophils, etc. In step S220, the target cell population of interest is selected in the FSC-SSC scatter plot by gating, such as... Figure 3 As shown.

[0033] For a selected target cell population, flow cytometry can generate a histogram. In step S230, negative and positive cell populations are selected from this histogram by gating, such as... Figure 4 As shown in the diagram. Positive cells correspond to cells stained with fluorescence, while negative cells correspond to cells that are not stained with fluorescence.

[0034] Furthermore, flow cytometers include multiple channels for receiving and detecting the fluorescence signals emitted by a sample upon excitation. Each channel corresponds to a different fluorescent dye, and each channel is used to detect its corresponding fluorescent dye. When a single-stained sample stained with a particular fluorescent dye is input, the strongest fluorescence signal can be detected in the channel corresponding to that specific fluorescent dye.

[0035] In step S240, based on the negative and positive cell populations selected in step S230, and based on the fluorescence signal detection results of the single-stained sample by flow cytometry, a scatter plot of the main channel relative to each of the multiple sub-channels is generated. The main channel is the channel corresponding to the j-th fluorescent dye, and the remaining channels are sub-channels. For example, Figure 5 A scatter plot illustrating the main channel relative to the first secondary channel is shown schematically. Figure 5 The diagram shows two separate cell populations: a positive cell population and a negative cell population. The straight line connecting the centers of these two populations is not horizontal, but has a certain slope.

[0036] In step S250, the slope S of the line is calculated using the following mathematical formula (1):

[0037]

[0038] It should be noted that the slope S mentioned above is calculated based on a scatter plot of the main channel relative to a subchannel (e.g., the first subchannel). Therefore, the slope can be calculated separately for each scatter plot of the generated main channel relative to each subchannel. In this paper, it is assumed that the flow cytometer includes L fluorescence channels, so L-1 slopes can be calculated.

[0039] Furthermore, this paper assumes the use of P fluorescent dyes, therefore P single-stained samples are sequentially input in step S210, and steps S210-S250 are repeated P times.

[0040] After performing steps S210-S250 on P single-stained samples stained with P fluorescent dyes, an initial overflow matrix can be generated based on the calculated slope, as shown in step S260. Specifically, the overflow matrix M is defined by the following mathematical formula (2):

[0041]

[0042] Where i represents the fluorescence channel of the flow cytometer, and i = 1, 2, ..., L. j represents the type of fluorescent dye, and j = 1, 2, ..., P. The slope S calculated according to mathematical formula (1) can be used as an element m of the overflow matrix M. ij Specifically, given a single-stained sample that has been stained with the j-th fluorescent dye, element m... ij It is equal to the slope value calculated based on the scatter plot of the determined main channel relative to the i-th channel (if the main channel is not the i-th channel), or equal to the value "100%" (if the main channel is the i-th channel).

[0043] The following will combine Figure 6 To describe in detailFigure 1 Steps S120 and S130 in the process. Figure 6 A flowchart is shown illustrating a method for obtaining unmixed results and analyzing the obtained unmixed results according to the present disclosure.

[0044] like Figure 6 As shown, in step S610, the flow cytometer input is... Figure 2 The single-staining sample used in step S210 that has been stained with the j-th dye is, in other words, the single-staining sample used when generating the initial overflow matrix is ​​still used at this time.

[0045] At this point, the signals detected by the detectors in each fluorescence channel of the flow cytometer can be represented as... Where j represents the j-th fluorescent dye used to stain the currently input single-stained sample, and L represents the number of fluorescent channels.

[0046] In step S620, the detected signal Y and the overflow matrix M represented by mathematical formula (2) are substituted into the following mathematical formula (3) to calculate the demixing result.

[0047] Y = MX - (3)

[0048] X represents the signal values ​​corresponding to the P fluorescence channels when a single-stained sample stained with the j-th dye is input.

[0049] In step S630, a scatter plot of the main channel relative to each of the multiple secondary channels is generated based on the demixing result (e.g., ...). Figure 7 (As shown). Here, the main channel is the channel corresponding to the j-th fluorescent dye among the P channels, and the other P-1 channels are secondary channels. Then, in step S640, it can be determined whether the unmixing result for the currently input single-stained sample meets the requirements based on these scatter plots.

[0050] Figure 7 The scatter plot of the main channel relative to two of the P-1 subchannels is shown only schematically. For example, in the scatter plot of the main channel relative to the 5th subchannel, the straight line connecting the centers of the positive and negative populations is horizontal, indicating that the negative population (cells not stained by the j-th dye) and the positive population (cells stained by the j-th dye) are identified only in the main channel, while there is no positive population corresponding to the j-th dye in the 5th subchannel. That is, the currently input single-stained sample has no effect on the 5th subchannel. In this case, the main channel and the 5th subchannel can be considered well demixed.

[0051] Furthermore, in the scatter plot of the main channel relative to the third sub-channel, the straight line connecting the centers of the positive and negative populations slopes significantly upwards, indicating that the negative and positive populations can also be roughly identified in the third sub-channel. In other words, the currently input monochromatic sample has an impact on the third sub-channel. In this case, it can be assumed that the main channel and the third sub-channel are not well unmixed.

[0052] However, the above is only a rather rough, intuitive analysis. To quantitatively analyze the unmixing results, more precise metrics are needed. This disclosure proposes using the slope of the straight line connecting the centers of the positive and negative populations as an indicator to determine whether the unmixing results meet the requirements. In the following text, this slope is also referred to as the compensation gap.

[0053] To calculate the slope, known methods can be used, such as linear regression (e.g., normal least squares) or robust linear regression (e.g., random sampling consensus algorithm (RANSAC)).

[0054] Furthermore, this disclosure provides a simpler method for calculating the slope. Specifically, as described above, the method for obtaining the unmixed result still uses the same single-stained sample used when generating the initial overflow matrix; therefore, the negative and positive cells in the sample remain unchanged. Therefore, when generating the scatter plot of the main channel relative to P-1 secondary channels in step S630, the sample can be directly used... Figure 2 The negative and positive cell populations selected in step S230 mean that it is not necessary to perform target cell population selection and negative and positive cell population selection on the single-stained sample input in step S610. To achieve this, in Figure 2 In step S230, the indexes of the selected negative and positive cells can be further stored. Thus, in step S630, the negative cell population and the positive cell population can be determined based on the stored indexes, thereby generating a scatter plot.

[0055] After generating the scatter plot (e.g.) Figure 7 As shown, the slope S of the straight line connecting the centers of two populations can be calculated for each scatter plot according to mathematical formula (1), which is the compensation gap. The compensation gap reflects the difference between the current unmixing result and the ideal result (the straight line is horizontal). For the current input monochromatic sample, when the compensation gaps calculated for P-1 scatter plots are all not greater than a predetermined threshold, the unmixing result is considered to meet the requirements. Conversely, when at least one of the calculated compensation gaps is greater than the predetermined threshold, the unmixing result is considered to have failed to meet the requirements. Those skilled in the art can set the predetermined threshold according to actual needs.

[0056] existFigure 6 In the flowchart shown, steps S610-S640 are performed on all P single-stained samples stained with P dyes. If the unmixing results for all P single-stained samples meet the requirements, it can be assumed that the generated initial overflow matrix will be able to effectively unmix the actual sample to be detected, because the actual sample is stained with multiple dyes, which is equivalent to a combination of different single-stained samples. In this case, the initial overflow matrix can be provided as the final overflow matrix for detecting the actual sample, such as... Figure 1 The steps are shown in step S140.

[0057] Conversely, if the analysis indicates that the obtained unmixing result does not meet the requirements, this disclosure provides a compensation matrix for correcting the initial overflow matrix.

[0058] The compensation matrix C is defined by the following mathematical expression (4):

[0059]

[0060] Where i represents the fluorescent channel, and i = 1, 2, ..., P. j represents the type of fluorescent dye, and j = 1, 2, ..., P. Given a single-stained sample stained with the j-th fluorescent dye, element c... ij It is equal to the compensation gap calculated based on the scatter plot of the main channel relative to the i-th channel (when the main channel is not the i-th channel), or equal to the value "100%" (when the main channel is the i-th channel).

[0061] Then, the modified overflow matrix M can be calculated according to the following mathematical formula (5). c :

[0062] M c =MC-(5)

[0063] Then, the modified overflow matrix M can be provided. c As the final overflow matrix used to detect actual samples, such as Figure 1 The steps are shown in step S140.

[0064] On the other hand, if the unmixing result does not meet the requirements, the method described in this disclosure can also notify the user to regenerate the initial overflow matrix (returning to...). Figure 1 Instead of correcting the initial overflow matrix as described above, step S110) is used. In the case of regenerating the initial overflow matrix, for example, the following measures can be taken:

[0065] - If the unmixing result is poor only for a single-stained sample stained with a certain dye, the single-stained sample can be remade with that dye.

[0066] - Change the dye and use a dye with low similarity to create a single-stained sample;

[0067] - Reselect the positive and negative cell populations.

[0068] It should be noted that the above are merely examples of some possible adjustments, and this disclosure is not limited thereto. Those skilled in the art can use other suitable known adjustment methods.

[0069] The following will combine Figures 8-10B Here is an example illustrating the user interface of a flow cytometer according to this disclosure. Users can... Figure 8 In the interface shown, adjust the parameters used to generate the initial overflow matrix, such as selecting the target cell population in the FSC-SSC scatter plot, and selecting the negative and positive populations in the histogram.

[0070] Users can Figure 9 View the demixing results in the interface shown. Figure 9 The upper chart 910 shows the spectral characteristics of various monostained samples after demixing, where the horizontal axis represents the fluorescence channels and the vertical axis represents the fluorescence intensity. Figure 9 The lower part shows the similarity matrix 920 and complexity 930, which users can use to analyze the unmixing results.

[0071] Furthermore, various algorithms can be employed when using the overflow matrix for demixing. Therefore, Figure 9 The interface also includes an algorithm switcher 940, which allows users to switch between different algorithms, making it easy for them to view and compare the unmixing effects when using various algorithms. For example... Figure 9 As illustrated, the algorithm switcher 940 indicates that the least squares method (LSM) is currently being used, and a "Change" button is provided on the right for changing the algorithm.

[0072] Users can Figure 10A and Figure 10B Viewed in the interface shown Figure 6 The scatter plot obtained in step S630. Figure 10A A scatter plot of the main channel relative to P-1 secondary channels obtained through step S630 is schematically shown. Figure 10B A scatter plot is shown schematically. The horizontal axis represents the primary channel (e.g., BUV737-A), and the vertical axis represents the secondary channel (e.g., BUV496-A, etc.).

[0073] Figure 10A and Figure 10BThe calculation of the compensation gap 1010 for the corresponding scatter plot is also shown, such as "0.0", "0.2", etc. When the calculated compensation gap is greater than a predetermined threshold, the value of the compensation gap is not displayed; instead, a warning indicator 1020 is displayed. The warning indicator 1020 can remind the user that the current unmixing result deviates significantly from the ideal result.

[0074] Therefore, users can clearly view the unmixing results for the current monochromated sample and easily notice poor unmixing results. Furthermore, although not shown in the accompanying drawings, the user interface according to this disclosure may also provide a controller for switching between various monochromated samples, facilitating users to view and compare the unmixing effects for different monochromated samples.

[0075] On the other hand, as the number of channels in flow cytometers increases—for example, some flow cytometers now contain as many as 88 channels—listing all channels on the user interface for selection or adjustment is no longer feasible. Listing all channels would consume significant screen space and make operation cumbersome. Furthermore, inexperienced users adjusting individual channels may affect spectral characteristics and lead to inaccurate results. To address these issues, this disclosure provides a new user interface, which will be discussed below. Figures 11A-13 Provide a detailed description.

[0076] See Figure 11A The interface shown allows users to set the commonly used FSC and BSSC channels in the first row. If users need to set the gain values ​​for other SSC channels, they can click the "Show channels" button 1110, which will then display the FSC channels and all SSC channels, as shown. Figure 11B As shown.

[0077] also, Figure 11A Gain adjusters 1120 for individual lasers are shown, allowing users to simultaneously adjust all fluorescence channels of a given laser by simply sliding a slider or directly specifying the desired gain percentage. In this case, the specific fluorescence channel is hidden to avoid affecting the spectral characteristics due to inexperienced users individually adjusting the gain value of each channel.

[0078] For users with sufficient expertise and experience, clicking the "Show channels" button 1130 on the right side of the corresponding laser will allow them to view all the fluorescence channels of the laser for configuration. Figure 11B The 20 fluorescence channels U1-U20 corresponding to the UV of the ultraviolet laser are schematically shown.

[0079] In addition, withFigure 11A The "Display Channel" buttons 1110 and 1130 in the middle correspond to each other. Figure 11B The interface also provides "Hide channels" buttons 1110' and 1130', which can be clicked to return to the previous screen. Figure 11A The interface shown.

[0080] If the user modifies the channel gain, or if the user-set gain value is a boundary value (e.g., "1" or "3000"), the corresponding channel and laser can be marked. For example, as Figure 11C As shown, the gain values ​​of channels U1 and U8 are modified, and the gain values ​​of channels U5 and U9 are the boundary values ​​"1" and "3000" respectively. Therefore, the names of channels U1, U5, U8, and U9, and the names of the corresponding lasers, are appended with the symbol "*". By doing so, users can easily identify the channels whose gain values ​​have been modified or whose gain values ​​have reached the boundary values, as well as their corresponding lasers.

[0081] Figure 12 The diagram schematically illustrates a user interface for setting channels on the axes of a plot such as an FSC-SSC scatter plot, where channels are grouped by laser, as indicated by the arrows in the figure. Furthermore, a hierarchical menu is used to display lasers and channels. For example, a parent menu displays various lasers (such as Violet, Blue, etc.), and a submenu corresponding to a specific laser displays multiple channels associated with that laser (such as V1-A, V2-A, etc.). This hierarchical menu simplifies channel display and avoids consuming excessive screen space. Moreover, users do not need to search through a long list of channels (e.g., a list of 88 channels) but can simply move within a small area and click to select the desired channel.

[0082] Figure 13 The diagram schematically illustrates the user interface for setting up channels on an oscilloscope, where channels are grouped by laser, as shown by the boxes in the figure. Furthermore, each laser and its corresponding channel can be displayed in a different color to distinguish them from one another. For example, a red laser and its corresponding channel can be displayed in red, and a yellow laser and its corresponding channel in yellow.

[0083] The technology according to this disclosure has been described above in conjunction with specific embodiments. This disclosure provides a compensation gap metric, enabling users to quantitatively analyze whether the unmixing results using the overflow matrix meet requirements. Furthermore, if the unmixing results do not meet requirements, this disclosure provides a compensation matrix composed of the compensation gap to correct the overflow matrix, eliminating the need for users to regenerate or manually adjust the overflow matrix. In addition, this disclosure provides a novel user interface on which users can easily view the quality of the unmixing results, conveniently view and select laser channels, and easily set the gain of the laser channels.

[0084] It should be noted that the methods described in this disclosure are not necessarily to be executed in the order shown in the flowchart. Where technically feasible, some steps in the methods may be executed in different orders or in parallel.

[0085] The information processing methods for flow cytometers described above can be implemented by software, hardware, or a combination of both. Programs included in the software can be stored beforehand in a storage medium located internally or externally to the device. As an example, during execution, these programs are written to random access memory (RAM) and executed by a processor (e.g., a CPU) to implement the various methods and processes described herein. Therefore, this disclosure also includes an information processing system for flow cytometers, comprising a processing unit configured to perform the methods described above.

[0086] In addition, this disclosure also includes computer program code and computer program products for implementing the methods described above, and computer-readable storage media on which the computer program code is recorded.

[0087] Figure 14 An example configuration block diagram of computer hardware is shown for performing the methods of this disclosure according to a program.

[0088] like Figure 14 As shown, in computer 1400, central processing unit (CPU) 1401, read-only memory (ROM) 1402 and random access memory (RAM) 1403 are connected to each other via bus 1404.

[0089] The input / output interface 1405 is further connected to the bus 1404. The input / output interface 1405 is connected to the following components: an input device 1406 formed by a keyboard, mouse, microphone, etc.; an output device 1407 formed by a display, speaker, etc.; a storage device 1408 formed by a hard disk, non-volatile memory, etc.; a communication device 1409 formed by a network interface card (such as a local area network (LAN) card, modem, etc.); and a driver 1410 for driving a removable medium 1411, such as a magnetic disk, optical disk, magneto-optical disk, or semiconductor memory.

[0090] In a computer with the above structure, the CPU 1401 loads a program stored in the storage device 1408 into the RAM 1403 via the input / output interface 1405 and the bus 1404, and executes the program to perform the method described above.

[0091] The program to be executed by the computer (CPU 1401) can be recorded on a removable medium 1411, which is formed as a packaging medium, such as a disk (including a floppy disk), an optical disk (including a compact optical disk-read-only memory (CD-ROM)), a digital multifunction optical disk (DVD), etc.), a magneto-optical disk, or a semiconductor memory. Furthermore, the program to be executed by the computer (CPU 1401) can also be provided via wired or wireless transmission media such as a local area network, the Internet, or digital satellite broadcasting.

[0092] When the removable medium 1411 is installed in the drive 1410, the program can be installed in the storage device 1408 via the input / output interface 1405. Alternatively, the program can be received by the communication device 1409 via a wired or wireless transmission medium and installed in the storage device 1408. Alternatively, the program can be pre-installed in the ROM 1402 or the storage device 1408.

[0093] The modules or systems described in this disclosure are for logical purposes only and do not strictly correspond to physical devices or entities. For example, the function of each module described in this disclosure may be implemented by multiple physical entities, or the function of multiple modules described in this disclosure may be implemented by a single physical entity. Furthermore, features, components, elements, steps, etc., described in one embodiment are not limited to that embodiment, but can also be applied to other embodiments, such as replacing specific features, components, elements, steps, etc., in other embodiments, or in combination with them.

[0094] The scope of this disclosure is not limited to the specific embodiments described herein. Those skilled in the art will understand that various modifications or variations can be made to the embodiments described herein, depending on design requirements and other factors, without departing from the principles of this disclosure. The scope of this disclosure is defined by the appended claims and their equivalents.

Claims

1. An information processing method for flow cytometers, comprising: Based on the detection results of multiple single-stained samples input to the flow cytometer, an initial overflow matrix is ​​generated, wherein each single-stained sample is stained with a corresponding one of a variety of fluorescent dyes; For each of the plurality of single-stained samples, The generated initial overflow matrix is ​​used to unmix the detection results for the single-stained sample; Based on the unmixing results, a first scatter plot of the main channel relative to each of the multiple secondary channels is generated, and the first slope of the straight line connecting the center of the positive cell population and the center of the negative cell population in each first scatter plot is calculated as a compensation gap. Based on multiple compensation gaps calculated for the multiple monostained samples, a final overflow matrix is ​​provided for detecting actual samples.

2. The information processing method according to claim 1 further includes: If none of the calculated compensation gaps are greater than a predetermined threshold, the initial overflow matrix is ​​provided as the final overflow matrix. If at least one of the plurality of compensation gaps is greater than the predetermined threshold, one of the following operations shall be performed: The initial overflow matrix is ​​corrected, and the corrected overflow matrix is ​​provided as the final overflow matrix; Regenerate the initial overflow matrix.

3. The information processing method according to claim 2 further includes: The initial overflow matrix is ​​corrected using a compensation matrix, wherein the compensation matrix is ​​constructed based on a plurality of calculated compensation gaps.

4. The information processing method according to claim 1, wherein, The step of generating the initial overflow matrix further includes: For each of the plurality of single-stained samples, Identify the target cell population in the single-stained sample; Identify the negative and positive cells in the target cell population and store the indices of the negative and positive cells; Based on the detection results of the single-stained samples, a second scatter plot of the main channel relative to each of the multiple secondary channels is generated, and a second slope of the straight line connecting the center of the positive cell population and the center of the negative cell population in each second scatter plot is calculated; the initial overflow matrix is ​​constructed based on the multiple second slopes calculated for the multiple single-stained samples.

5. The information processing method according to claim 4 further includes: The negative cells and the positive cells are determined based on the stored index; The first scatter plot of the main channel relative to each sub-channel is generated based on the identified negative and positive cells.

6. The information processing method according to claim 1 or 4, wherein, Calculate each of the first slope and the second slope using the following mathematical formula:

7. The information processing method according to claim 1, further comprising: The display shows the result of demixing using the initial overflow matrix. This displays the controller used to switch between various demixing algorithms.

8. The information processing method according to claim 1, further comprising: The display shows the first scatter plot of the main channel relative to each secondary channel, and the compensation gap calculated based on the first scatter plot. Specifically, if the compensation gap is greater than the predetermined threshold, a warning indicator is displayed on the corresponding first scatter plot.

9. The information processing method according to claim 1, further comprising at least one of the following: The controller for adjusting the gain of all channels corresponding to one laser of the flow cytometer is displayed, while the controller for adjusting the gain of each channel individually is hidden. Displays a mark used to identify the following channels and their corresponding lasers: the gain of the channel has been adjusted, or the gain of the channel has reached a boundary value; Multiple lasers are displayed in the parent menu, and multiple channels corresponding to one of the lasers are displayed in the sub-menu corresponding to that laser. Each laser and its corresponding channel are displayed in different colors.

10. An information processing system for a flow cytometer, comprising a processing unit configured to: Based on the detection results of multiple single-stained samples input to the flow cytometer, an initial overflow matrix is ​​generated, wherein, Each single-stained sample is stained with one of a variety of fluorescent dyes; For each of the plurality of single-stained samples, The generated initial overflow matrix is ​​used to unmix the detection results for the single-stained sample; Based on the unmixing results, a first scatter plot of the main channel relative to each of the multiple secondary channels is generated, and the first slope of the straight line connecting the center of the positive cell population and the center of the negative cell population in each first scatter plot is calculated as a compensation gap. Based on multiple compensation gaps calculated for the multiple monostained samples, a final overflow matrix is ​​provided for detecting actual samples.

11. The information processing system according to claim 10, wherein, The processing unit is further configured to: If none of the calculated compensation gaps are greater than a predetermined threshold, the initial overflow matrix is ​​provided as the final overflow matrix. If at least one of the plurality of compensation gaps is greater than the predetermined threshold, one of the following operations shall be performed: The initial overflow matrix is ​​corrected, and the corrected overflow matrix is ​​provided as the final overflow matrix; Regenerate the initial overflow matrix.

12. The information processing system according to claim 11, wherein, The processing unit is also configured to correct the initial overflow matrix using a compensation matrix, wherein the compensation matrix is ​​constructed based on a plurality of calculated compensation gaps.

13. The information processing system according to claim 10, wherein, The processing unit is also configured to perform the following operation to generate the initial overflow matrix: For each of the plurality of single-stained samples, Identify the target cell population in the single-stained sample; Identify the negative and positive cells in the target cell population and store the indices of the negative and positive cells; Based on the detection results of the single-stained samples, a second scatter plot of the main channel relative to each of the multiple secondary channels is generated, and a second slope of the straight line connecting the center of the positive cell population and the center of the negative cell population in each second scatter plot is calculated; the initial overflow matrix is ​​constructed based on the multiple second slopes calculated for the multiple single-stained samples.

14. The information processing system according to claim 13, wherein, The processing unit is further configured to: The negative cells and the positive cells are determined based on the stored index; The first scatter plot of the main channel relative to each sub-channel is generated based on the identified negative and positive cells.

15. The information processing system according to claim 10 or 13, wherein, The processing unit is also configured to calculate each of the first slope and the second slope according to the following mathematical formula:

16. The information processing system according to claim 10, wherein, The processing unit is also configured to perform at least one of the following operations on the user interface of the flow cytometer: The display shows the result of demixing using the initial overflow matrix. This displays the controller used to switch between various demixing algorithms. The system displays the first scatter plot of the main channel relative to each secondary channel and the compensation gap calculated for the first scatter plot, wherein if the compensation gap is greater than the predetermined threshold, a warning indicator is displayed for the corresponding first scatter plot.

17. The information processing system according to claim 10, wherein, The processing unit is also configured to perform at least one of the following operations on the user interface of the flow cytometer: The controller for adjusting the gain of all channels corresponding to one laser of the flow cytometer is displayed, while the controller for adjusting the gain of each channel individually is hidden. Displays a mark used to identify the following channels and their corresponding lasers: the gain of the channel has been adjusted, or the gain of the channel has reached a boundary value; Multiple lasers are displayed in the parent menu, and multiple channels corresponding to one of the lasers are displayed in the sub-menu corresponding to that laser. Each laser and its corresponding channel are displayed in different colors.

18. A flow cytometer comprising an information processing system according to any one of claims 10 to 17.

19. A non-transitory computer-readable storage medium storing instructions that, when executed by a processor, cause the processor to perform the information processing method according to any one of claims 1 to 9.

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