Test method and test device for flow cytometer, and flow cytometer

Abstract

The invention discloses a testing method and a testing device for a flow cytometer and the flow cytometer. A flow cytometer includes a plurality of lasers arranged collinearly and emitting light of different wavelengths. The testing method for the flow cytometer comprises the following steps: performing a first group of tests, and in the first group of tests, adopting a first testing reagent as a sample, sequentially turning on a plurality of lasers one by one and turning off the other lasers to test the fluorescence channel performance of each of the plurality of lasers.

Description

Technical Field

[0001] The present disclosure relates to the technical field of flow cytometric detection, and in particular to a testing method and a testing device for a flow cytometer, and a flow cytometer. Background Art

[0002] This section provides background information related to the present disclosure which is not necessarily prior art.

[0003] In flow cytometers using collinear laser designs, crosstalk occurs between fluorescence signals excited by different lasers, interfering with the performance of the original signal. For example, a 405nm violet laser has a higher energy density than a 488nm blue laser. The fluorescence signal generated by the 405nm laser will dominate the fluorescence signal generated by the 488nm laser. The resulting signal result for the 488nm fluorescence channel may not represent the true performance of that channel. Therefore, a more suitable method for testing the performance of fluorescence channels in such flow cytometers is needed. Summary of the Invention

[0004] A brief overview of the present disclosure is provided below to provide a basic understanding of certain aspects of the present disclosure. However, it should be understood that this overview is not an exhaustive overview of the present disclosure. It is not intended to identify key or important parts of the present disclosure, nor is it intended to limit the scope of the present disclosure. Its purpose is simply to present certain concepts of the present disclosure in a simplified form as a prelude to the more detailed description that will be given later.

[0005] In view of the above, an object of the present disclosure is to provide an improved testing method, testing device, and flow cytometer for a flow cytometer, which eliminate the crosstalk between fluorescence signals excited by different lasers.

[0006] According to one aspect of the present disclosure, a testing method for a flow cytometer is provided. The flow cytometer includes a plurality of lasers arranged collinearly and emitting light of different wavelengths. The testing method includes performing a first set of tests. In the first set of tests, a first test reagent is used as a sample, and the plurality of lasers are sequentially enabled one by one while the remaining lasers are disabled to test the fluorescence channel performance of each of the plurality of lasers.

[0007] According to another aspect of the present disclosure, a testing device for a flow cytometer is provided. The flow cytometer includes a plurality of lasers arranged collinearly and emitting light of different wavelengths. The testing device includes a processing circuit configured to: perform a first set of tests in which a first test reagent is used as a sample, sequentially turning on the plurality of lasers one by one and disabling the remaining lasers to test fluorescence channel performance of each of the plurality of lasers.

[0008] According to yet another aspect of the present disclosure, a flow cytometer comprising the above-mentioned testing device is provided.

[0009] According to other aspects of the present disclosure, a computer program code and a computer program product for implementing the above-mentioned testing method according to the present disclosure, as well as a computer-readable storage medium having recorded thereon the computer program code for implementing the above-mentioned testing method according to the present disclosure, are also provided.

[0010] Other aspects of the embodiments of the present disclosure are given in the following description, wherein the detailed description is used to fully disclose the preferred embodiments of the embodiments of the present disclosure without imposing limitations thereon. BRIEF DESCRIPTION OF THE DRAWINGS

[0011] The present disclosure may be better understood by referring to the detailed description given below in conjunction with the accompanying drawings, in which the same or similar reference numerals are used throughout the drawings to represent the same or similar parts. The accompanying drawings, together with the following detailed description, are incorporated into and form a part of this specification and are used to further illustrate the preferred embodiments of the present disclosure and to explain the principles and advantages of the present disclosure. Among them:

[0012] Figure 1 is a flowchart illustrating an example of a process of a testing method for a flow cytometer according to an embodiment of the present disclosure;

[0013] Figure 2 is an example simplified structural diagram of a flow cytometer;

[0014] Figures 3A to 3D is a schematic diagram illustrating a test method for a flow cytometer in conjunction with an example simplified structure of the flow cytometer;

[0015] Figure 4 is a flowchart of an example process of another testing method for a flow cytometer according to an embodiment of the present disclosure;

[0016] Figure 5 is a schematic diagram of a test report obtained using the test method according to an embodiment of the present disclosure;

[0017] Figure 6 is a block diagram illustrating a configuration example of a test apparatus for flow cytometer according to an embodiment of the present disclosure; and

[0018] Figure 7 FIG. 4 is a block diagram of an exemplary structure of a general-purpose personal computer in which the testing method according to an embodiment of the present disclosure can be implemented. DETAILED DESCRIPTION

[0019] Exemplary embodiments of the present disclosure are described below with reference to the accompanying drawings. For the sake of clarity and conciseness, not all features of an actual implementation are described in this specification. However, it should be understood that in the process of developing any such actual implementation, many implementation-specific decisions must be made in order to achieve the developer's specific goals, such as compliance with system and business-related constraints, which may vary from implementation to implementation. In addition, it should be understood that although the development work may be very complex and time-consuming, it is a routine task for those skilled in the art who benefit from the contents of this disclosure.

[0020] It should be understood that although the terms "first," "second," etc. may be used herein to describe various elements, these elements should not be limited by these terms. These terms are only used to distinguish one element from another. For example, a first element may be referred to as a second element, and similarly, a second element may be referred to as a first element, without departing from the scope of the present disclosure.

[0021] It is also necessary to explain here that, in order to avoid obscuring the present disclosure due to unnecessary details, the accompanying drawings only show the device structure and / or processing steps closely related to the scheme according to the present disclosure, while other details that are not closely related to the present disclosure are omitted.

[0022] The embodiments of the present disclosure are described in detail below with reference to the accompanying drawings.

[0023] First, combine Figure 1 、 Figure 2 as well as Figures 3A to 3D An implementation example of a testing method for a flow cytometer according to an embodiment of the present disclosure is described below. Figure 1 is a flowchart illustrating an example of a flow of a testing method for a flow cytometer according to an embodiment of the present disclosure. Figure 2 is an example simplified block diagram of a flow cytometer, and Figures 3A to 3D The diagram illustrates a test method for a flow cytometer by using an example simplified structure of the flow cytometer.

[0024] like Figure 1 As shown, the testing method according to an embodiment of the present disclosure may begin at step S110 and end at step S120. The testing method according to an embodiment of the present disclosure may be used in a quality control process. The quality control process is intended to verify the quality of a flow cytometer, and key functions and performance of the cytometer are typically checked during this process.

[0025] In step S110, multiple lasers are turned on one by one and the remaining lasers are disabled. Figure 2 and Figures 3A to 3DThis is explained in more detail.

[0026] like Figure 2 As shown, the flow cytometer 100 includes a plurality of lasers arranged in a collinear manner and emitting light of different wavelengths, such as a first laser 111, a second laser 112, a third laser 113 and a fourth laser 114. It should be noted that, in order not to confuse the key points, Figure 2 Only some of the components of the flow cytometer 100 are shown. The flow cytometer 100 may also include many components in addition to the components shown. For example, a beam expander, a lens module, etc. may be included between each laser and the corresponding dichroic mirror. For example, a lens module, etc. may be included between the fourth dichroic mirror 124 and the flow cell module 130. In addition, the four lasers shown are only examples, and the flow cytometer 100 may include a greater or lesser number of lasers. As an example, the first laser 111 may emit a 638nm laser, the second laser 112 may emit a 561nm laser, the third laser 113 may emit a 488nm laser, and the fourth laser 114 may emit a 405nm laser. Of course, the present disclosure is not limited to this.

[0027] exist Figure 2 In the flow cytometer, light emitted from different lasers is reflected by corresponding dichroic mirrors (first dichroic mirror 121, second dichroic mirror 122, third dichroic mirror 123, and fourth dichroic mirror 124), and the light path is guided to the flow cytometer module 130. In the flow cytometer module 130, cells in the sample scatter the light generated by the lasers, and the fluorescein on the cells is excited by the lasers. The generated light signals (e.g., fluorescent signals, non-fluorescent scattered signals such as side scatter signals and forward scatter signals) are collected and processed by the detection system 140.

[0028] In a flow cytometer designed with spatially separated lasers, all lasers are turned on during the quality control process. Since different lasers illuminate the cells in the sample using different light paths, the generated fluorescence signals will not have crosstalk. However, in a flow cytometer 100 designed with collinear lasers, such as Figure 2 As shown in FIG, when all lasers are turned on at the same time, since the lasers generated by different lasers illuminate the cells in the sample through the same optical path, crosstalk will occur between the fluorescence signals excited by different lasers, and the crosstalk signal will interfere with the performance of the original signal.

[0029] Therefore, when testing the fluorescence channel performance, in step S110, one of the multiple lasers is turned on at a time, and the other lasers are turned off. Then, in step S120, the fluorescence channel performance at the laser wavelength emitted by the laser is tested. In this way, multiple lasers are turned on one by one until the fluorescence channel performance of all lasers is tested. Figure 3AIn the embodiment, only the first laser 111 is turned on to test the fluorescence channel performance of the first laser 111. Figure 3B In the embodiment, only the second laser 112 is turned on to test the fluorescence channel performance of the second laser 112. Figure 3C In the embodiment, only the third laser 113 is turned on to test the fluorescence channel performance of the third laser 113. Figure 3D In the embodiment, only the fourth laser 114 is turned on, so as to test the fluorescence channel performance of the fourth laser 114 .

[0030] Specifically, testing the performance of the fluorescence channel may include: after turning on a certain laser among the multiple lasers, adjusting the gain of the signal of the fluorescence channel so that the median of the fluorescence signal is the target median; and determining whether the median of the fluorescence signal after gain adjustment is within a predetermined range, and / or determining whether the coefficient of variation (CV) or the robust coefficient of variation (rCV) is less than a predetermined value, and / or determining whether the deviation between the gain after adjustment and the gain before adjustment is within a predetermined range. Here, determining whether the median is within the predetermined range is a further confirmation of the gain adjustment result. As an example, when these three conditions are met at the same time, it can be determined that the performance of the fluorescence channel meets the requirements.

[0031] Thus, the testing method according to the embodiments of the present application can eliminate crosstalk between different sensors in a collinear laser design, thereby accurately understanding the fluorescence channel performance of each laser. Furthermore, because the fluorescence channel performance of each laser can be accurately measured and controlled within predetermined requirements, the quality control process can ensure that the fluorescence channel performance of the flow cytometer is well suited for testing actual samples.

[0032] The following will be combined Figure 4 Let's discuss a process example of another testing method for a flow cytometer according to an embodiment of the present application. Figure 4 FIG. 4 is a flowchart of another exemplary process of a testing method for a flow cytometer according to an embodiment of the present disclosure. Figure 1 The test method in Figure 4 Corresponding to test S209.

[0033] First, the user clicks the quality control button on the flow cytometer to initiate the quality control process, which then proceeds to test S201. In test S201, the laser power is tested. Test S201 may include simultaneously turning on each laser, reading the power value of each laser, and determining whether the power value of each laser is within the corresponding target power range.

[0034] Next, in test S202, the sheath fluid flow rate is tested. It should be noted that the flow cytometer according to the present application may specifically include a flow sensor to test the sheath fluid flow rate. Test S202 may include: reading the sheath fluid flow rate via the flow sensor, and then determining whether the sheath fluid flow rate is within a target rate range.

[0035] A stable sheath fluid flow rate can ensure the stability and standard deviation of the flow cytometer's system signal. In particular, in nano-flow cytometers used to detect extremely small particles such as nanoparticles or extracellular vesicles, the sheath fluid flow rate can also affect the system's sensitivity to small particle signals. Therefore, by measuring the sheath fluid flow rate and controlling it within a predetermined range, the stability of the system signal can be ensured, and the system's sensitivity to small particle signals can be improved. Of course, for traditional flow cytometers (used for detecting larger particle sizes), the sheath fluid flow rate can also be tested during its quality control process to ensure the stability of the system signal.

[0036] Next, in test S203, background noise is measured. In the present application, the background noise measured includes at least one of photoelectric threshold, sheath fluid noise, and sample noise. Photoelectric threshold refers to noise caused by the photoelectric element, sheath fluid noise refers to noise caused by the sheath fluid (e.g., particles and impurities in the sheath fluid), and sample noise refers to noise caused by the sample line (e.g., particles and impurities in the sample line).

[0037] For example, the photoelectric threshold can be obtained by measuring the system noise without both sheath fluid and sample flowing, the sheath fluid noise can be obtained with only sheath fluid flowing without sample flowing, and the sample noise can be obtained with both sheath fluid and sample flowing.

[0038] As an example, the test S203 may include: loading a sample, testing a photoelectric threshold, sheath fluid noise, and sample noise; and determining whether the photoelectric threshold, sheath fluid noise, and sample noise are within respective target ranges.

[0039] It should be noted that in this application, when testing background noise, high-purity water or internal sheath fluid is used as the sample. In particular, when using internal sheath fluid as the sample, there is no need to prepare a separate liquid as the sample, which can simplify the testing process and reduce testing costs.

[0040] By controlling various background noise levels within a predetermined range, system sensitivity for small particle signals can be improved. This is particularly true in nano-flow cytometers used to detect extremely small particles, such as nanoparticles and extracellular vesicles, which are extremely sensitive to background noise. Background noise can affect system sensitivity for small particle signals, a key characteristic of the system.

[0041] Next, the first and second sets of tests will be described. Because it is more important to understand the performance of the side scatter channel for small signal levels, the two sets of tests use different test reagents. The first test reagent used in the first set of tests contains a fluorescent dye, while the second test reagent used in the second set of tests has smaller test beads than the first test reagent and does not contain a fluorescent dye. For example, the first test reagent is a 500nm test bead with a single peak, and the second test reagent is a 144nm polystyrene (PS) bead.

[0042] Prior to test S204, the previous sample is unloaded, the sample line is cleaned, and the second test reagent is loaded. In test S204, it is tested whether the second test reagent is loaded. Test S204 may include: testing the number of events per second (EPS); and determining whether the EPS is greater than a predetermined value.

[0043] Next, in tests S205 and S206 , the side scatter channel performance is tested.

[0044] Test S205 may include: adjusting the gain of the side scatter channel signal so that the median of the side scatter signal reaches a target median; and determining whether the median of the side scatter signal after the gain adjustment is within a predetermined range, and / or determining whether a deviation between the gain after the adjustment and the gain before the adjustment is within a predetermined range. For example, if both conditions are met, it can be determined that the performance of the side scatter channel meets the requirements.

[0045] Test S206 may include: testing the sensitivity of the side scatter channel, where sensitivity is represented by the degree of separation between signal and noise; and determining whether the sensitivity is within a predetermined range. For example, if all conditions in tests S205 and S206 are simultaneously met, the side scatter channel performance may be determined to meet the requirements.

[0046] The inventors of this application discovered through experiments that when a sample fluid flows through a flow cytometer, the side scatter signal has two peaks, and the number of side scatter signal peaks does not change with the number of particle types in the sample fluid. Through analysis, the inventors found that the peak with the greater signal intensity of the two side scatter signal peaks corresponds to the fluorescent signal emitted by the sample fluid, while the peak with the smaller signal intensity (i.e., the peak with the smallest signal intensity) corresponds to background noise. Therefore, by determining the distance between the first and second subsets of the sample signal corresponding to the two peaks (the first peak and the second peak), respectively, the sensitivity of the side scatter channel can be determined.

[0047] As an example, the distance between the first subset and the second subset can be represented by Fisher distance FD. In this case, the above distance can be obtained by the following formula (1):

[0048]

[0049] In formula (1), MFI1 and MFI2 represent the medians of the first subset and the second subset, respectively, and δ1 and δ2 represent the standard deviations of the first subset and the second subset, respectively.

[0050] Therefore, the sensitivity of the side scatter channel can be expressed by FD.

[0051] Next, before test S207, the previous sample is unloaded, the sample line is cleaned, and the first test reagent is loaded. In test S207, it is tested whether the first test reagent is loaded. Test S207 may include: testing the number of events per second (EPS); and determining whether the EPS is greater than a predetermined value.

[0052] Next, in test S208 , the delay between the multiple lasers is tested. The delay between the multiple lasers is caused by the time difference between the particles in the sample being irradiated by the lasers emitted by different lasers.

[0053] The test S208 may include: testing the delay between the plurality of lasers; and determining whether the delay is within a predetermined range.

[0054] Next, in test S209 , the plurality of lasers are turned on one by one in sequence and the remaining lasers are disabled to test the fluorescence channel performance of each of the plurality of lasers.

[0055] Test S209 may include: after turning on a laser from among the multiple lasers, adjusting the gain of the fluorescence channel signal so that the median of the fluorescence signal is a target median; determining whether the median of the fluorescence signal after the gain adjustment is within a predetermined range; and / or determining whether the coefficient of variation (CV) or the robust coefficient of variation (rCV) is less than a predetermined value; and / or determining whether the deviation between the gain after adjustment and the gain before adjustment is within a predetermined range. For example, if all three conditions are met simultaneously, the fluorescence channel performance may be determined to meet the requirements.

[0056] Next, in test S2010, the forward scatter channel performance is tested. Test S2010 may include: adjusting the gain of the forward scatter channel signal so that the median of the forward scatter signal is a target median; and determining whether the median of the forward scatter signal after gain adjustment is within a predetermined range, and / or determining whether the coefficient of variation or robust coefficient of variation is less than a predetermined value, and / or determining whether the deviation between the gain after adjustment and the gain before adjustment is within a predetermined range. As an example, if both conditions are met simultaneously, the forward scatter channel performance can be determined to meet the requirements.

[0057] It should be noted that Figure 4The test method in the embodiment is only a possible embodiment, wherein the tests S204, S205 and S206 need to be performed after the second test reagent is loaded, and the tests S207, S208, S209 and S2010 need to be performed after the first test reagent is loaded. Figure 4 For example, the order of tests S208, S209 and S2010 in the first group of tests can be swapped, the order of tests S205 and S206 in the second group of tests can be swapped, and the order of tests S201, S202, S203, the first group of tests and the second group of tests can also be swapped. In addition, the order of tests S201, S202, S203, the first group of tests and the second group of tests can also be swapped. Figure 4 Part of the test.

[0058] Therefore, by dividing the tests related to the fluorescence channel, the forward scattering channel, and the side scattering channel into two groups, the performance of different channels can be tested more specifically, and better evaluation results of the performance of different channels can be obtained.

[0059] In addition, a test report (eg, a quality control report) may be generated after all tests are completed or when some tests fail to present the test results to the user. Figure 5 is a schematic diagram of a test report obtained using the test method according to an embodiment of the present disclosure.

[0060] like Figure 5 As shown, the test report can include various parameters obtained during the above-mentioned test process, such as the test results of the laser (delay, default delay, delay difference, power, target power, etc.), sheath fluid flow rate (test value, target value, etc.), background noise (test value and target value of photoelectric threshold, sheath fluid noise, sample noise, etc.), side scatter channel performance (EPS, gain, target gain, deviation from target gain, median, target median, deviation from target median, FD, target FD, etc.), fluorescence channel performance and forward scatter channel performance (EPS, gain, target gain, deviation from target gain, median, target median, deviation from target median, rCV, target rCV, etc.), test requirements, quality test results (i.e., pass or fail), etc.

[0061] exist Figure 5 In the test example shown, some of the test requirements are as follows:

[0062] In the performance test of the laser, -4.00μs ≤ delay difference (the difference between the delay and the default delay) ≤ 4.00μs,

[0063] In the performance tests of the side scatter channel, forward scatter channel, and fluorescence channel, -20.00% ≤ the deviation (%) of the adjusted gain from the target gain ≤ 20.00%, -5.00% ≤ the deviation (%) of the adjusted median from the target median ≤ 5.00%, where the target gain is the gain before gain adjustment and is the system default gain when the system is first started.

[0064] In the test of the performance of the fluorescence channel, rCV (%) ≤ target rCV (%),

[0065] In the performance test of the side scatter channel, FD ≥ target FD.

[0066] also, Figure 5 Below the channel column in the chart are the labels of different channels, where the VFSC channel is the test result of the forward scattering channel.

[0067] The quality control report allows users to understand the current status of the flow cytometer at a glance.

[0068] The above description has described a test method for a flow cytometer according to an embodiment of the present disclosure. Corresponding to the above embodiment of the test method for a flow cytometer, the present disclosure further provides the following embodiment of a test device for a flow cytometer. Figure 6 is a block diagram illustrating a configuration example of a test apparatus 200 for flow cytometry according to an embodiment of the present disclosure.

[0069] like Figure 6 As shown, the test device 200 according to an embodiment of the present disclosure may include a processing circuit 210. The processing circuit 210 may be configured to: execute Figure 1 The test method shown or Figure 4 Tests S201-S2010 are shown.

[0070] In addition, according to an embodiment of the present disclosure, a flow cytometer including the above-mentioned testing device 200 may be provided. For example, the flow cytometer may include a nano-flow cytometer for detecting extremely small particles such as nanoparticles or extracellular vesicles, but is not limited thereto.

[0071] It should be noted that although the above describes the test method and test device for a flow cytometer and the functional configuration and operation of the flow cytometer according to the embodiments of the present disclosure, this is only an example and not a limitation, and those skilled in the art may modify the above embodiments according to the principles of the present disclosure, for example, the functional modules and operations in each embodiment may be added, deleted or combined, and such modifications shall fall within the scope of the present disclosure.

[0072] In addition, it should be pointed out that the device embodiment here corresponds to the above-mentioned method embodiment. Therefore, for the content not described in detail in the device embodiment, please refer to the description of the corresponding part of the method embodiment, and will not be repeated here.

[0073] In addition, the present disclosure also provides a storage medium and a program product. It should be understood that the machine-executable instructions in the storage medium and program product according to the embodiments of the present disclosure can also be configured to perform the above-mentioned test method. Therefore, for any content not described in detail herein, reference can be made to the description of the corresponding parts previously described, and the description will not be repeated here.

[0074] Accordingly, the storage medium for carrying the program product including the machine executable instructions is also included in the disclosure of the present invention. The storage medium includes but is not limited to a floppy disk, an optical disk, a magneto-optical disk, a memory card, a memory stick, and the like.

[0075] In addition, it should be noted that the above series of processes and devices can also be implemented by software and / or firmware. In the case of implementation by software and / or firmware, the data is transmitted from a storage medium or a network to a computer with a dedicated hardware structure, such as Figure 7 The general-purpose personal computer 1300 shown is installed with programs constituting the software. When various programs are installed, the computer can execute various functions and the like.

[0076] exist Figure 7 In the embodiment, a central processing unit (CPU) 1301 executes various processes according to a program stored in a read-only memory (ROM) 1302 or a program loaded from a storage device 1308 to a random access memory (RAM) 1303. In the RAM 1303, data required when the CPU 1301 executes various processes and the like is also stored as needed.

[0077] The CPU 1301, the ROM 1302, and the RAM 1303 are connected to one another via a bus 1304. An input / output interface 1305 is also connected to the bus 1304.

[0078] The following components are connected to the input / output interface 1305: an input device 1306 including a keyboard, a mouse, etc.; an output device 1307 including a display such as a cathode ray tube (CRT), a liquid crystal display (LCD), etc., and a speaker, etc.; a storage device 1308 including a hard disk, etc.; and a communication device 1309 including a network interface card such as a LAN card, a modem, etc. The communication device 1309 performs communication processing via a network such as the Internet.

[0079] A drive 1310 is also connected to the input / output interface 1305 as needed. A removable medium 1311 such as a magnetic disk, an optical disk, a magneto-optical disk, a semiconductor memory, etc. is mounted on the drive 1310 as needed so that a computer program read therefrom is installed in the storage device 1308 as needed.

[0080] In the case of realizing the above-described series of processing by software, a program constituting the software is installed from a network such as the Internet or a storage medium such as the removable medium 1311 .

[0081] It should be understood by those skilled in the art that such storage media is not limited to Figure 7 The removable medium 1311 shown has a program stored therein and is distributed separately from the device to provide the program to the user. Examples of removable medium 1311 include magnetic disks (including floppy disks (registered trademark)), optical disks (including compact disk read-only memories (CD-ROMs) and digital versatile disks (DVDs)), magneto-optical disks (including minidiscs (MDs) (registered trademark)), and semiconductor memories. Alternatively, the storage medium may be ROM 1302, a hard disk included in storage device 1308, or the like, in which the program is stored and distributed to the user along with the device containing it.

[0082] The preferred embodiments of the present disclosure are described above with reference to the accompanying drawings, but the present disclosure is of course not limited to the above examples. Those skilled in the art may obtain various changes and modifications within the scope of the appended claims, and it should be understood that these changes and modifications will naturally fall within the technical scope of the present disclosure.

[0083] For example, a plurality of functions included in one unit in the above embodiments may be implemented by separate devices. Alternatively, a plurality of functions implemented by a plurality of units in the above embodiments may be implemented by separate devices, respectively. In addition, one of the above functions may be implemented by a plurality of units. Needless to say, such a configuration is included in the technical scope of the present disclosure.

[0084] In this specification, the steps described in the flowchart include not only processing executed in time series in the order described, but also processing executed in parallel or individually rather than necessarily in time series. In addition, even in the steps processed in time series, it goes without saying that the order can be changed as appropriate.

[0085] The various techniques described in this specification can be performed independently of each other unless a conflict arises. Of course, any of the various techniques can be performed in combination. In one example, part or all of the techniques described in any embodiment can be performed in combination with part or all of the techniques described in another embodiment. In addition, any part or all of the techniques described above can be performed in combination with another technique not described above.

Claims

1. A test method for a flow cytometer comprising a plurality of lasers arranged collinearly and emitting light of different wavelengths, the test method comprising performing a first set of tests, In the first set of tests, the first test reagent is used as a sample, the plurality of lasers are turned on one by one in sequence and the remaining lasers are disabled to test the fluorescence channel performance of each of the plurality of lasers.

2. The testing method according to claim 1, wherein: Testing the fluorescence channel performance includes: After turning on a certain laser among the plurality of lasers, adjusting the gain of the signal of the fluorescence channel so that the median of the fluorescence signal is the first target median; and Determine whether the median of the fluorescence signal after gain adjustment is within a first predetermined range, and / or determine whether the coefficient of variation is less than a first predetermined value, and / or determine whether the deviation between the gain after adjustment and the gain before adjustment is within a second predetermined range.

3. The testing method according to claim 1, wherein: The first set of tests also includes testing delays between the plurality of lasers.

4. The testing method according to claim 1, wherein: The first set of tests also includes: testing the forward scatter channel performance, Wherein, testing the forward scattering channel performance includes: Adjusting the gain of the signal of the forward scatter channel so that the median of the forward scatter signal is the second target median; and Determine whether the median of the forward scattered signal after the gain is adjusted is within a third predetermined range, and / or determine whether the coefficient of variation is less than a second predetermined value, and / or determine whether the deviation between the gain after the adjustment and the gain before the adjustment is within a fourth predetermined range.

5. The testing method according to claim 1, wherein: The testing method further includes: performing a second set of tests, the second set of tests using a second test reagent as a sample, the second test reagent having a smaller test bead than the first test reagent and not containing a fluorescent dye, and The second set of tests at least includes: testing side scatter channel performance.

6. The testing method according to claim 5, wherein: Testing the side scatter channel performance includes: adjusting the gain of the signal of the side scatter channel so that the median of the side scatter signal is the third target median; and It is determined whether the median of the side scatter signal after the gain adjustment is within a fifth predetermined range, and / or whether a deviation between the gain after the adjustment and the gain before the adjustment is within a sixth predetermined range.

7. The testing method according to claim 6, wherein: Testing the side scatter channel performance also includes: The side scatter channel was tested for sensitivity, which was expressed as the separation of signal from noise.

8. The testing method according to claim 1, further comprising: The background noise is tested, where the background noise includes at least one of a photoelectric threshold, sheath fluid noise, and sample noise.

9. The testing method according to claim 8, wherein: When testing the background noise, high-purity water or internal sheath fluid is used as a sample.

10. The testing method according to claim 1, wherein: The testing method further comprises: testing the sheath fluid flow rate by using a flow sensor included in the flow cytometer. 11 . A testing device for a flow cytometer, the testing device comprising a processing circuit, the processing circuit being configured to: perform the testing method according to any one of claims 1 to 10.

12. A flow cytometer comprising the testing device according to claim 11. 13 . A computer-readable storage medium storing instructions, which, when executed by a processor, cause the processor to perform the testing method according to claim 1 .

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