Information processing apparatus, particle analyzing apparatus, particle sorting apparatus, and information processing method

Abstract

To provide a technique capable of improving reliability of data analysis.SOLUTION: The present technology includes a storage unit that stores event data including light intensity data obtained by irradiating one particle of a plurality of particles with light, and a processing unit that performs processing on a plurality of pieces of event data acquired from the plurality of particles, wherein the storage unit stores a flag that is given when the light intensity data exceeds a threshold value, the processing unit performs processing on the plurality of pieces of event data other than the light intensity data to which the flag is assigned, in response to an instruction to exclude the light intensity data to which the flag is assigned.SELECTED DRAWING: Figure 1

Description

[Technical Field]

[0001] The present technology relates to an information processing device, a particle analysis device, a particle sorting device, and an information processing method, and more particularly to an information processing device, a particle analysis device, a particle sorting device, and an information processing method that can improve the reliability of data analysis. [Background technology]

[0002] Conventionally, a device (e.g., a flow cytometer) has been used to measure the individual characteristics of particles by labeling particles such as cells with a fluorescent dye and irradiating them with laser light to detect the fluorescence and scattered light emitted from the particles. In such a device, the light reaching the photodetector is converted into an electrical signal (voltage pulse) and digitized. Statistical analysis, etc., is then performed based on the numerical data for each parameter.

[0003] In recent years, multicolor measurements have been performed in which particles are labeled with multiple fluorescent dyes and the light emitted from each fluorescent dye is measured using multiple photodetectors with different wavelength bands. However, in multicolor measurements, fluorescence from fluorescent dyes other than the target dye can leak into each photodetector. To address this issue and improve the reliability of data analysis, fluorescence compensation is performed, which subtracts the leaked fluorescence intensity from the fluorescence intensity measured by the photodetector. Fluorescence compensation involves applying electrical or mathematical corrections to the pulses in a dedicated circuit so that the fluorescence intensity measured by the photodetector corresponds to the true fluorescence intensity from the target fluorescent dye.

[0004] For example, Patent Document 1 discloses a method for calculating the true fluorescence intensity from a target fluorescent dye by expressing the fluorescence intensity measured by each photodetector as a vector and applying the inverse matrix of a preset leakage matrix to this vector.

[0005] In contrast to this, Patent Document 2 discloses a method for calculating true fluorescent dyes from each fluorescent dye by approximating the measured spectrum by a linear sum of single staining spectra, without using the inverse matrix of the leakage matrix. [Prior art documents] [Patent documents]

[0006] [Patent Document 1] Japanese Patent Application Laid-Open No. 2003-83894 [Patent Document 2] Japanese Patent Application Laid-Open No. 2011-232259 Summary of the Invention [Problem to be solved by the invention]

[0007] However, with conventional instruments, when signals from photodetectors are converted from analog to digital, if the signal level is high enough to exceed the upper detection limit of the photodetector, not only will the input signal data to that photodetector be inaccurate, but the signal will also be affected by signals obtained from other photodetectors during fluorescence compensation processing, etc., which can lead to problems that reduce the reliability of the overall measurement data. In such cases, it becomes necessary to adjust the gain during data collection, and in the data analysis stage it becomes necessary to create gates to exclude the maximum values ​​in the plots of each parameter axis from the measured data.

[0008] Therefore, the main object of this technology is to provide a technology that can improve the reliability of data analysis. [Means for solving the problem]

[0009] That is, the present technology provides an information processing device that includes a memory unit that stores event data consisting of light intensity data obtained by irradiating light on one of a plurality of particles, and a processing unit that processes the plurality of event data acquired from the plurality of particles, wherein the memory unit stores a flag that is assigned when the light intensity data exceeds a threshold, and the processing unit processes the plurality of event data other than the light intensity data that has been assigned the flag in response to an instruction to exclude the light intensity data that has been assigned the flag.

[0010] The present technology also provides a particle analysis device including a light irradiation unit that irradiates light onto one particle out of a plurality of particles, a light detection unit that detects light from the one particle, a memory unit that stores event data including light intensity data obtained from the light detection unit, and a processing unit that processes a plurality of event data acquired from the plurality of particles, wherein the memory unit stores a flag that is assigned when the light intensity data exceeds a threshold, and the processing unit processes the plurality of event data other than the light intensity data that has been assigned the flag in response to an instruction to exclude the light intensity data to which the flag has been assigned.

[0011] Furthermore, the present technology also provides a particle sorting device comprising: a light irradiation unit that irradiates light onto one particle of a plurality of particles; a light detection unit that detects light from the one particle; a memory unit that stores event data consisting of light intensity data obtained from the light detection unit; and a processing unit that processes the plurality of event data acquired from the plurality of particles, wherein the memory unit stores a flag that is assigned when the light intensity data exceeds a threshold, and the processing unit processes the plurality of event data other than the light intensity data to which the flag is assigned in response to an instruction to exclude the light intensity data to which the flag is assigned, and further comprises a sorting unit that sorts particles linked to the plurality of event data other than the event data including the light intensity data in response to an instruction to exclude the light intensity data to which the flag is assigned.

[0012] In addition, the present technology also provides an information processing method including a storage step of storing event data consisting of light intensity data obtained by irradiating light on one of a plurality of particles, and a processing step of processing the plurality of event data acquired from the plurality of particles, wherein the storage step stores a flag that is assigned when the light intensity data exceeds a threshold, and the processing step processes the plurality of event data other than the light intensity data that has been assigned the flag in accordance with an instruction to exclude the light intensity data that has been assigned the flag. [Brief explanation of the drawings]

[0013] [Figure 1] 1 is a schematic conceptual diagram illustrating an example of an embodiment of an information processing device according to the present technology. [Figure 2] 1A is a plot showing event data that does not include light intensity data that exceeds the threshold, and FIG. 1B is a plot showing event data that includes light intensity data that exceeds the threshold. [Figure 3] FIG. 1A is a plot diagram showing the case where gating is performed on event data that does not contain light intensity data exceeding the threshold, and FIG. 1B is a plot diagram showing the case where gating is performed on event data that contains light intensity data exceeding the threshold. [Figure 4] 1A to 1C are diagrams showing examples of plot diagram displays. [Figure 5] FIG. 10 is a diagram showing an example of the ratio of event data including flagged light intensity data being output and displayed on a display unit. [Figure 6] 10 is a flowchart illustrating a first processing example. [Figure 7] 10A and 10B are diagrams showing examples of data display in processing example 1 and processing example 2. [Figure 8] 10 is a flowchart illustrating a second processing example. [Figure 9] 10 is a flowchart illustrating a processing example 3. [Figure 10] 10 is a flowchart illustrating a processing example 4. [Figure 11] 10 is a flowchart illustrating a processing example 5. [Figure 12] 1 is a schematic conceptual diagram illustrating an example of an embodiment of a particle analysis device according to the present technology. [Figure 13] 1 is a schematic conceptual diagram showing an example of an embodiment of a particle sorting device according to the present technology. [Figure 14] FIG. 10 is a schematic conceptual diagram showing another example of an embodiment of a particle sorting device according to the present technology. DETAILED DESCRIPTION OF THE INVENTION

[0014] A preferred embodiment for carrying out the present technology will be described below. The embodiments described below are representative embodiments of the present technology, and should not be construed as narrowing the scope of the present technology. The present technology will be described in the following order. 1. First embodiment (information processing device) (1) Storage section 11 (2) Processing section 12 (3) User Interface 13 (4)Display section 14 (5) Processing example by the processing unit 12 <Processing example 1> <Processing example 2> <Processing example 3> <Processing example 4> <Processing example 5> 2. Second embodiment (particle analysis device) (1) Light irradiation unit 21 (2) Photodetector 22 (3) Processing section 12 3. Third embodiment (particle sorting device) (1) Preparation section 31 4. Fourth embodiment (information processing method)

[0015] 1. First embodiment (information processing device)

[0016] 1 is a schematic conceptual diagram showing a first embodiment. An information processing device 10 according to this embodiment includes a storage unit 11 and a processing unit 12. If necessary, the information processing device 10 may also include other units such as a user interface 13 and a display unit 14.

[0017] (1) Storage section 11

[0018] The storage unit 11 stores event data consisting of light intensity data obtained by irradiating light onto one of the particles, and also stores a flag that is assigned when the light intensity data exceeds a threshold value.

[0019] In the present technology, the "particle" particularly refers to a microparticle, and the microparticle can be appropriately selected. In the present technology, the microparticle can include, for example, biological microparticles such as cells, cell aggregates, microorganisms, and ribosomes, as well as synthetic microparticles such as gel particles, beads, latex particles, polymer particles, and industrial particles. Biological microparticles (also referred to as "bioparticles") may include chromosomes, ribosomes, mitochondria, organelles, and the like that constitute various cells. Cells may include animal cells (e.g., blood cells) and plant cells. The cells may be, in particular, blood cells or tissue cells. The blood cells may be, for example, suspension cells such as T cells and B cells. The tissue cells may be, for example, adherent cultured cells or adherent cells dissociated from tissue. Cell aggregates may include, for example, spheroids and organoids. Microorganisms may include bacteria such as Escherichia coli, viruses such as tobacco mosaic virus, and fungi such as yeast. Furthermore, the biological microparticles may also include biological macromolecules such as nucleic acids, proteins, and complexes thereof. These biological macromolecules may be, for example, extracted from cells or contained in a blood sample or other liquid sample. The synthetic microparticles may be, for example, microparticles made of organic or inorganic polymeric materials or metals. Organic polymeric materials may include polystyrene, styrene-divinylbenzene, polymethyl methacrylate, etc. Inorganic polymeric materials may include glass, silica, magnetic materials, etc. Metals may include gold colloids, aluminum, etc. The synthetic microparticles may be, for example, gel particles, beads, etc., and in particular, gel particles or beads to which one or a combination of two or more selected from oligonucleotides, peptides, proteins, and enzymes is bound.

[0020] The shape of the particles may be spherical or nearly spherical, or may be non-spherical. The size and mass of the particles may be selected as appropriate. In this technology, chemical or biological labels, such as fluorescent dyes and fluorescent proteins, may be attached to the particles as needed. The labels to be attached may be selected as appropriate. Molecules that specifically react with the particles (e.g., antibodies, aptamers, DNA, RNA, etc.) may be bound to the labels. In the present technology, the particles are preferably biological particles, and may particularly be cells.

[0021] The fluorescent dye for labeling the particles is not particularly limited, and at least one of known dyes used for staining biological particles, etc. can be used. For example, fluorescent dyes include phycoerythrin (PE), fluorescein isothiocyanate (FITC), PE-Cy5, PE-Cy7, PE-TexasRed (registered trademark), allophycocyanin (APC), APC-Cy7, ethidium bromide, and propidium iodide. iodide), Hoechst (registered trademark) 33258, Hoechst (registered trademark) 33342, DAPI (4',6-diamidino-2-phenylindole), acridine orange, chromomycin, mithramycin, olivomycin, pyronin Y, thiazole orange, rhodamine 101, isothiocyanate, BCECF, BCECF-AM, C.SNARF-1, C.SNARF-1-AMA, aequorin, Indo-1, Indo-1-AM, Fluo-3, Examples of fluorescent dyes that can be used include Fluo-3-AM, Fura-2, Fura-2-AM, oxonol, Texas Red (registered trademark), rhodamine 123, 10-N-nony-acridine orange, fluorescein, fluorescein diacetate, carboxyfluorescein, carboxyfluorescein diacetate, carboxydichlorofluorescein, and carboxydichlorofluorescein diacetate. Derivatives of the above-mentioned fluorescent dyes can also be used.

[0022] In this embodiment, light intensity data is generated by receiving, with a photodetector, fluorescence or scattered light generated by irradiating light onto particles, and event data is generated based on the light intensity data. In addition, when particles are labeled with a fluorescent dye, light intensity data is generated by irradiating light onto the fluorescent dye-labeled particles and receiving, with a photodetector, fluorescence generated from the excited fluorescent dye.

[0023] More specifically, when a photodetector receives fluorescence, scattered light, or the like, the corresponding electrical signal output from the photodetector is input to an analog-to-digital converter (ADC). This ADC is a circuit downstream (output side) of the photodetector and is connected to the photodetector. The electrical signal is an analog signal obtained by photoelectrically converting the light detected by the photodetector. Each ADC then converts each input electrical signal from analog to digital. Each ADC then outputs the converted digital electrical signal to the downstream stage.

[0024] The electrical signals output from each analog-digital conversion circuit are input to the data detection circuit. This data detection circuit is a circuit subsequent to (on the output side of) each analog-digital conversion circuit and is connected to each analog-digital conversion circuit. The data detection circuit uses a specific signal from each input electrical signal as a trigger signal for particle detection. That is, when the value of the trigger signal satisfies a predetermined condition, the data detection circuit detects that each electrical signal has been detected from a particle. The data detection circuit then reads the waveform of each input electrical signal and calculates the parameters (width, height, and area) of the read waveform to generate light intensity data. Furthermore, the data detection circuit generates event data associated with a corresponding particle based on each light intensity data, which is the calculated parameter value of the waveform. In this embodiment, the memory unit 11 stores the light intensity data and event data generated in this manner.

[0025] In the present technology, as described above, the storage unit 11 stores a flag to be assigned when the light intensity data exceeds a threshold. Specifically, when the storage unit 11 receives a signal indicating that the light intensity data exceeds a threshold, the storage unit 11 associates information indicating whether each piece of light intensity data exceeds the threshold with each piece of light intensity data and stores the information based on the signal. More specifically, the storage unit 11 assigns the flag to light intensity data that exceeds the threshold and stores the information.

[0026] The flag may be set, for example, when the light from the particle is detected by the photodetector and the upper limit of the detection voltage of the photodetector is exceeded. More specifically, for example, when the signal from the photodetector is converted into analog digital data by the analog digital conversion circuit, the input voltage range of the analog digital conversion is exceeded, or when the upper limit of the data that can be held is exceeded when the digital signal is processed. When processing a digital signal, if the upper limit of the data that can be held is exceeded, specifically, if the upper limit of the data that can be held, i.e., the upper limit of the value that can be expressed in bit width, is exceeded, it is necessary to perform processing to cut off (clip) the excess portion. During this clipping processing, the above flag can be assigned.

[0027] The flag may be set, for example, when the light intensity data exceeds an upper limit of data processing when it is processed. More specifically, for example, when the light intensity data exceeds an upper limit of data that can be held when generating event data based on the light intensity data, or when the light intensity data or event data exceeds an upper limit of data that can be held when converted into a transfer data format when transferring the light intensity data or event data from an information processing device according to the present technology to a data analysis device (for example, a personal computer, a server, etc.).

[0028] In this embodiment, the event data may be composed of a plurality of light intensity data obtained by irradiating the particle with a plurality of light beams. In this case, the plurality of light beams may be emitted from a plurality of light sources that are configured to irradiate excitation light beams of different wavelengths. In this embodiment, for example, the plurality of light intensity data may be obtained by irradiating light beams from the plurality of light sources at different positions and detecting the light beams from the particle with different photodetectors.

[0029] (2) Processing section 12

[0030] The processing unit 12 processes the plurality of event data acquired from the plurality of particles. Furthermore, in response to an instruction to exclude the flagged light intensity data, the processing unit 12 processes the plurality of event data other than the flagged light intensity data.

[0031] In conventional devices, when a signal from a photodetector is subjected to analog-to-digital conversion and light intensity data exceeding the threshold is detected, there is no means for excluding the light intensity data and processing the data. Figure 2A is a plot diagram showing event data that does not include light intensity data exceeding the threshold, and Figure 2B is a plot diagram showing event data that includes light intensity data exceeding the threshold. As such, in conventional devices, it is not possible to exclude event data that includes light intensity data exceeding the threshold, and therefore the only option during the data measurement stage was to adjust the gain so that the threshold is not exceeded.

[0032] 3A is a plot diagram when gating is performed on event data that does not contain light intensity data exceeding the threshold, and FIG. 3B is a plot diagram when gating is performed on event data that does contain light intensity data exceeding the threshold. In this way, with conventional devices, when event data containing light intensity data exceeding the threshold is present in the data analysis stage, the only option is to perform analysis while allowing some light intensity data exceeding the threshold to be mixed in, for example, by creating a gate that excludes the maximum value data in the plot of each parameter axis for the measured data.

[0033] Furthermore, if a photodetector detects light intensity data exceeding the threshold, not only will the input signal data for that photodetector be inaccurate, but it will also affect the signals obtained by other photodetectors in fluorescence compensation processing, etc. As a result, there is a problem of reduced reliability in data analysis.

[0034] In contrast, in the present technology, the processing unit 12 can process the plurality of event data other than the light intensity data to which the flag has been assigned in response to an instruction to exclude the light intensity data to which the flag has been assigned. Therefore, when displaying, analyzing, or processing the obtained event data, the processing unit 12 has a means for excluding the light intensity data to which the flag has been assigned, and as necessary, data that may undermine the reliability of the results can be excluded from the measured data, thereby improving the reliability of the data analysis.

[0035] More specifically, when generating information for performing fluorescence correction such as unmixing and compensation (e.g., spectral reference, compensation matrix, etc.), it is possible to determine whether or not to exclude light intensity data to which the flag is attached, and to use the data generated based on this determination, thereby obtaining more reliable data. This processing will be described later in "(5) Example of processing by processing unit 12."

[0036] In this embodiment, when the event data consists of a plurality of light intensity data obtained by irradiating a single particle with a plurality of lights, the processing unit 12 can process the plurality of event data other than the event data including the light intensity data in response to an instruction to exclude the light intensity data to which the flag is assigned.

[0037] In this case, the processing unit 12 may include an arithmetic processing unit 121 and an output processing unit 122. The arithmetic processing unit 121 calculates the proportion of event data that includes light intensity data to which the flag has been assigned. Furthermore, the output processing unit 122 performs processing to output the proportion of event data that includes light intensity data to which the flag has been assigned and / or processing to output a plot diagram.

[0038] In this embodiment, the arithmetic processing unit 121 may calculate the proportion of event data containing flagged light intensity data relative to event data (all event data) consisting of a series of multiple light intensity data, obtained by, for example, continuously irradiating multiple light beams onto each of multiple particles. Alternatively, the arithmetic processing unit 121 may calculate the proportion of event data containing flagged light intensity data relative to multiple event data included in a gated region on a plot diagram output by the output processing unit 122, which will be described later. Calculating these proportions can provide an index for a user to determine whether or not to exclude flagged light intensity data from all event data or multiple event data included in a gated region on the plot diagram.

[0039] In this embodiment, the output processing unit 122 can output, for example, the proportion of event data including light intensity data to which the flag is attached among a plurality of event data acquired from a plurality of particles to be analyzed. By outputting the proportion, for example, the user can use it as an index when determining whether or not to exclude the light intensity data to which the flag is attached.

[0040] Furthermore, the output processing unit 122 can output the proportion of event data including the flagged light intensity data to the display unit 14, which will be described later. Fig. 5 is a diagram showing an example of the proportion of event data including the flagged light intensity data being output and displayed on the display unit 14. In this case, the user can refer to the proportion and input via the user interface 13 whether or not to exclude the flagged light intensity data.

[0041] In addition, the output processing unit 122 may output a warning to the user when the ratio of event data including the flagged light intensity data exceeds a threshold. In the present technology, the threshold may be appropriately set by the user. The warning may be displayed on the display unit 14 described later, or may be notified to the user by sounding a sound, etc.

[0042] In this embodiment, the output processing unit 122 can output a plot created for a plurality of event data acquired from a plurality of particles to be analyzed. In this case, the above-mentioned calculation processing unit 121 further calculates the ratio of event data including light intensity data to which the flag is assigned to a plurality of event data included in a gated region on the plot.

[0043] Here, the proportion of event data containing flagged light intensity data may be output by the output processing unit 122 in a state where it is displayed on a plot diagram, or the proportion of event data alone may be output on a separate screen. When displayed on a plot diagram, it is also possible to display the proportion of event data on a newly created child plot diagram for multiple event data included in the gated region. This output allows the user to evaluate the reliability of the event data that the user wishes to analyze in detail, enabling more accurate analysis.

[0044] In this embodiment, the output processing unit 122 may output a warning to the user when the ratio of event data including the flagged light intensity data to a plurality of event data included in the gated region on the plot exceeds a threshold. In this technology, the threshold may be appropriately set by the user. The warning may be displayed on the display unit 14 (described later) or may be notified to the user by sound, etc.

[0045] In this embodiment, in response to an instruction to exclude the flagged light intensity data, the processing unit 12 can perform a sorting process on particles associated with the plurality of event data other than the event data including the flagged light intensity data. This allows only particles with highly reliable data to be selected and sorted. This process will be described later in "(1) Sorting unit 31."

[0046] Furthermore, in this embodiment, the processing unit 12 can perform processing to switch the display method of the spectral plot. A spectral plot is a plot diagram in which light intensity data obtained by irradiating a single light beam is displayed for each wavelength range of a photodetector, and can be displayed in different colors according to the frequency with which event data including predetermined light intensity data is detected, for example. Furthermore, when multiple beams of light are irradiated onto a particle, it is also possible to display a ribbon plot in which spectral plots corresponding to each irradiated beam are arranged (see FIG. 4). More specifically, the processing to switch the display method switches, for example, from a method in which all light intensity data obtained by analysis is displayed in different colors according to frequency to a method in which data with a frequency below a certain threshold is not displayed.

[0047] Figure 4A shows a plot diagram in which all event data is displayed in a color-coded manner according to frequency, Figure 4B shows a plot diagram in which a threshold is set and frequency data below that threshold is not displayed, and Figure 4C shows a plot diagram in which a threshold higher than that of Figure 4B is set and frequency data below that threshold is not displayed. By switching the display method in this way, it is possible to make the fluorescence spectral waveform of the main population (e.g., a single or multiple cell populations) more visible.

[0048] In this embodiment, the threshold value may be changed by the user as appropriate, or a fixed threshold value may be set in advance. When the threshold value is changed by the user as appropriate, the user may input the threshold value, or the threshold value may be selected using a slider.

[0049] (3) User Interface 13

[0050] The user inputs through the user interface 13. The user can access and control each unit of the information processing device according to this embodiment via the user interface 13. Note that the user interface 13 is not essential to the present technology, and an external operating device or the like may be connected. For example, a mouse, a keyboard, or the like can be used as the user interface 13.

[0051] In this embodiment, the instruction to exclude the flagged light intensity data can be executed by a user input via the user interface 13. The instruction to exclude the flagged light intensity data can be appropriately given via the user interface 13, regardless of the ratio of event data including the flagged light intensity data by the output processing unit 122 or the timing of outputting a warning based on that ratio. For example, the instruction can be appropriately given by selecting a button or the like displayed on the screen.

[0052] (4)Display section 14

[0053] The display unit 14 can display all information related to the analysis generated or output from each part of the device, such as information output by the processing unit 12. Note that the display unit 14 is not essential to the present technology, and an external display device or the like may be connected. For example, a display, a printer, or the like can be used as the display unit 14.

[0054] (5) Processing example by the processing unit 12

[0055] Hereinafter, examples of processing by the processing unit 12 will be described with reference to the respective flowcharts.

[0056] <Processing example 1>

[0057] FIG. 6 is a flowchart illustrating processing example 1. Processing example 1 is processing performed when registering a spectral reference. This spectral reference consists of a single-stain spectrum, which is used, for example, in unmixing processing. The single-stain spectrum is the fluorescence wavelength distribution of each fluorescent dye, and is light intensity data obtained by irradiating light onto particles individually labeled with each fluorescent dye, and receiving the fluorescence emitted from the excited fluorescent dye with a photodetector.

[0058] Unmixing is a fluorescence correction method in which particles labeled with multiple fluorescent dyes are measured by multicolor measurement, and the resulting measured spectrum is approximated by the linear sum of single-stained spectra using the weighted least squares method (WLSM) to obtain true light intensity data from each fluorescent dye. By performing unmixing processing, the spectral information of overlapping fluorescent dyes can be separated into each fluorescent dye, enabling highly accurate and reproducible analysis of fluorescent reagents and fluorescent proteins with similar fluorescent wavelength peaks.

[0059] First, the processing unit 12 receives the light intensity data to which the flag has been assigned (S1). In this case, the light intensity data consists of the single-stain spectrum described above. Next, the arithmetic processing unit 121 calculates the proportion of event data containing flagged light intensity data relative to all event data, which consists of a series of multiple light intensity data, obtained, for example, by continuously irradiating multiple light beams onto each of multiple particles. The output processing unit 122 then outputs the proportion (S2). If the proportion exceeds a threshold (S3), the output processing unit 122 issues a warning to the user (S4). Next, a determination is made as to whether or not to exclude the flagged light intensity data (S5). This determination may be made by the user, who inputs an instruction via the user interface 13.

[0060] If it is determined that the flagged light intensity data should be excluded (S5), the processing unit 12 excludes the flagged light intensity data (S6) and displays the data (S7). The data display method is not particularly limited, and the data may be displayed on the display unit 14 as a plot (including one-dimensional, two-dimensional, and three-dimensional), a spectrogram, a histogram (including one-parameter, two-parameter (cytogram, dot plot), and three-parameter), or the like. Specifically, the output processing unit 122 outputs a plot created for the plurality of event data other than the event data containing the flagged light intensity data. Figures 7A and 7B are diagrams showing examples of data display in processing example 1 and processing example 2 described below. Figure 7A is a plot with the vertical axis representing light intensity and the horizontal axis representing the photodetector channel, and Figure 7B is a histogram with the vertical axis representing the number of events and the horizontal axis representing the light intensity at the fluorescence peak wavelength at which the particle is labeled. On the other hand, if it is determined that the flagged intensity data should not be excluded (S5), the processing unit 12 displays the data as described above without excluding the flagged intensity data (S7).

[0061] Next, the displayed data is gated into positive and negative groups (S8). This gating process may be performed by the user, who inputs instructions via the user interface 13. Next, the processing unit 12 acquires the average light intensity values ​​for each of the positive and negative groups and performs differential processing (S9). Finally, the data obtained as a result of the differential processing is registered in the storage unit 11 as a spectral reference (S10).

[0062] <Processing example 2>

[0063] 8 is a flowchart illustrating processing example 2. Processing example 2 is a process for registering in a spectral reference, similar to processing example 1.

[0064] First, the processing unit 12 receives the light intensity data to which the flag has been assigned (S11). Note that in this case, the light intensity data consists of the single staining spectrum described above. Next, the processing unit 12 displays the data (S12). The data display method is as described above. In this case, the output processing unit 122 may output, for example, a plot created for the plurality of event data. Next, the displayed data is gated into positive and negative groups, respectively (S13). Note that this gating process may be performed by the user, in which case the user inputs instructions via the user interface 13.

[0065] Next, the calculation processing unit 121 calculates the proportion of event data containing the flagged light intensity data to the plurality of event data included in the gated region on the plot output by the output processing unit 122, and the output processing unit 122 outputs the proportion (S14). At this time, if the proportion exceeds a threshold (S15), the output processing unit 122 outputs a warning to the user (S16). Next, it is determined whether or not to exclude the flagged light intensity data (S17). Note that this determination may be made by the user, who inputs an instruction via the user interface 13.

[0066] If it is determined that the data should be excluded (S17), the light intensity data to which the flag is assigned is excluded (S18). Next, the processing unit 12 acquires the average light intensity value for each of the positive and negative groups and performs difference processing (S19). Finally, the data obtained as a result of the difference processing is registered in the storage unit 11 as a spectral reference (S20). On the other hand, if it is determined that the data should not be excluded (S17), the processing unit 12 performs the difference processing described above without excluding the flagged intensity data (S19), and registers the obtained data in the storage unit 11 as a spectral reference (S20).

[0067] <Processing example 3>

[0068] FIG. 9 is a flowchart illustrating Process Example 3. Process Example 3 is a process for performing automatic compensation. Compensation refers to the correction of fluorescence leakage. For example, in the case of two-color measurement, when two or more types of fluorescence are measured and their fluorescence wavelengths overlap, compensation is a fluorescence compensation method that electrically or mathematically corrects the amount of leakage between them. Mathematical fluorescence compensation methods include, for example, expressing the fluorescence intensity measured by each photodetector as a vector and applying the inverse matrix of a preset leakage matrix to this vector to calculate the true fluorescence intensity from the target fluorescent dye. This leakage matrix is ​​created by analyzing the single-stain spectrum described above, and is a column vector in which the fluorescence wavelength distribution of each fluorescent dye is arranged.

[0069] First, the processing unit 12 receives the light intensity data to which the flag has been assigned (S21). In this case, the light intensity data consists of the single-stain spectrum described above. Next, the arithmetic processing unit 121 calculates the proportion of event data containing flagged light intensity data relative to event data (all event data) consisting of a series of multiple light intensity data obtained by continuously irradiating multiple light beams onto each of multiple particles, and the output processing unit 122 outputs the proportion (S22). If the proportion exceeds a threshold (S23), the output processing unit 122 outputs a warning to the user (S24). Next, a determination is made as to whether or not to exclude the flagged light intensity data (S25). This determination may be made by the user, who inputs an instruction via the user interface 13.

[0070] If it is determined that the flagged light intensity data should be excluded (S25), the flagged light intensity data is excluded (S26). Next, the processing unit 12 displays the data in response to the instruction to exclude the flagged light intensity data (S27). The data display method is not particularly limited, and the data may be displayed on the display unit 14 as a plot (including one-dimensional, two-dimensional, and three-dimensional), a spectrogram, a histogram (including one-parameter, two-parameter (cytogram, dot plot), and three-parameter), or the like. In this case, the output processing unit 122 may output, for example, a plot created for the plurality of event data other than the event data including the flagged light intensity data. On the other hand, if it is determined that the flagged intensity data should not be excluded (S25), the processing unit 12 displays the data as described above without excluding the flagged intensity data (S27).

[0071] Next, the displayed data is gated to the positive or negative group (S28). The gating process may be performed by the user, who inputs instructions via the user interface 13. Finally, a matrix calculation is performed using the data obtained as a result of the gating process (S29).

[0072] <Processing example 4>

[0073] 10 is a flowchart illustrating processing example 4. Processing example 4 is a process for automatically performing compensation, similar to processing example 3.

[0074] First, the processing unit 12 receives the light intensity data to which the flag has been assigned (S31). Note that in this case, the light intensity data consists of the single staining spectrum described above. Next, the processing unit 12 displays the data (S32). The data display method is as described above. In this case, the output processing unit 122 may output, for example, a plot created for the plurality of event data. Next, the displayed data is gated into positive and negative groups, respectively (S33). Note that this gating process may be performed by the user, in which case the user inputs instructions via the user interface 13.

[0075] Next, the calculation processing unit 121 calculates the proportion of event data containing the flagged light intensity data to the plurality of event data included in the gated region on the plot output by the output processing unit 122, and the output processing unit 122 outputs the proportion (S34). At this time, if the proportion exceeds a threshold (S35), the output processing unit 122 outputs a warning to the user (S36). Next, it is determined whether or not to exclude the flagged light intensity data (S37). Note that this determination may be made by the user, who inputs an instruction via the user interface 13.

[0076] If it is determined that the flagged light intensity data should be excluded (S37), the flagged light intensity data is excluded (S38). Finally, a matrix calculation is performed using the obtained data (S39). On the other hand, if it is determined that the flagged intensity data should not be excluded (S37), the processing unit 12 performs a matrix calculation using the obtained data (S39) without excluding the flagged intensity data.

[0077] <Processing example 5> 11 is a flowchart illustrating processing example 5. Processing example 5 is processing when compensation is manually performed by the user.

[0078] First, the processing unit 12 receives the light intensity data to which the flag has been assigned (S41). The light intensity data in this case consists of a measured spectrum. The measured spectrum is light intensity data obtained by irradiating light onto particles multiply labeled with multiple fluorescent dyes having overlapping fluorescence wavelength bands, and receiving, with a photodetector, fluorescence emitted from the excited fluorescent dyes. Next, the arithmetic processing unit 121 calculates the proportion of event data containing flagged light intensity data relative to event data (all event data) consisting of a series of multiple light intensity data obtained by continuously measuring multiple particles, and the output processing unit 122 outputs the proportion (S42). If the proportion exceeds a threshold (S43), the output processing unit 122 outputs a warning to the user (S44). Next, a determination is made as to whether or not to exclude the flagged light intensity data (S45). The determination may be made by the user, who inputs an instruction via the user interface 13.

[0079] If it is determined that the flagged light intensity data should be excluded (S45), the processing unit 12 excludes the flagged light intensity data (S46). Next, the processing unit 12 displays the data in response to the instruction to exclude the flagged light intensity data (S47). The data display method is not particularly limited, and the data may be displayed on the display unit 14 as a plot (including one-dimensional, two-dimensional, and three-dimensional), a spectrogram, a histogram (including one-parameter, two-parameter (cytogram, dot plot), and three-parameter), or the like. In this case, the output processing unit 122 may output, for example, a plot created for the plurality of event data other than the event data including the flagged light intensity data. On the other hand, if it is determined that the flagged intensity data should not be excluded (S45), the processing unit 12 displays the data as described above without excluding the flagged intensity data (S47). Finally, the user visually corrects the displayed data (S48).

[0080] 2. Second embodiment (particle analysis device)

[0081] 12 is a schematic conceptual diagram showing a second embodiment. A particle analysis device 20 according to this embodiment comprises a light irradiation unit 21, a light detection unit 22, a memory unit 11, and a processing unit 12. Furthermore, as necessary, other units such as a user interface 13 and a display unit 14 may be included. In this embodiment, the memory unit 11, the user interface 13, and the display unit 14 are the same as those described above, and therefore a description thereof will be omitted here.

[0082] (1) Light irradiation unit 21

[0083] The light irradiation unit 21 irradiates one particle of the plurality of particles with light (e.g., excitation light, etc.). The light irradiation unit 21 may include, for example, a light source that emits light and an objective lens that focuses the excitation light on the particle. The light source may be appropriately selected by a person skilled in the art and may be, for example, a laser diode, an SHG laser, a solid-state laser, a gas laser, or a high-brightness LED, or a combination of two or more of these. Furthermore, the light irradiation unit 21 may include other optical elements as necessary in addition to the light source and the objective lens. For example, the light irradiation unit 21 may irradiate light at one position in the optical detection area, or may irradiate light at each of multiple positions.

[0084] In this embodiment, the light irradiating section 21 may have a plurality of light sources so as to be able to irradiate excitation light of different wavelengths.

[0085] (2) Photodetector 22

[0086] The light detection unit 22 detects light from the particle. More specifically, the light detection unit 22 detects scattered light and / or fluorescence generated from the particle upon irradiation by the light irradiation unit 21. The light detection unit 21 may include, for example, a condenser lens that condenses the fluorescence and / or scattered light generated from the particle and a photodetector. The photodetector may be, but is not limited to, a PMT, a photodiode, a CCD, a CMOS, or the like. In addition to the condenser lens and the detector, the light detection unit 22 may include other optical elements as needed. The light detection unit 22 may further include, for example, a spectroscopic unit. Examples of optical components constituting the spectroscopic unit include a grating, a prism, and an optical filter. The spectroscopic unit can, for example, detect light of a wavelength to be detected by separating it from light of other wavelengths.

[0087] The fluorescence detected by the light detection unit 22 may be, but is not limited to, fluorescence generated from the particle itself and fluorescence generated from a substance (e.g., a fluorescent substance) labeled on the microparticle. The scattered light detected by the light detection unit 22 may be forward scattered light, side scattered light, Rayleigh scattering, Mie scattering, or a combination thereof.

[0088] (3) Processing section 12

[0089] In this embodiment, the processing unit 12 can perform the following processes in addition to the processes described above. The processing unit 12 is connected to the light detection unit 22 and can analyze optical information detected by the light detection unit 22. More specifically, the processing unit 12 calculates feature quantities indicating the size, shape, internal structure, etc. of each particle from, for example, optical information of light received from the light detection unit 22 (for example, detected values ​​of fluorescence or scattered light, etc.).

[0090] In addition, in the present technology, an external analysis device or the like may be used for the analysis. Specifically, for example, the analysis may be performed by a personal computer or a CPU, or may be stored as a program in hardware resources including a recording medium (for example, a non-volatile memory (USB memory), HDD, CD, etc.) and operated by the personal computer or the CPU. Furthermore, the external analysis device or the like may be connected to each part of the device via a network.

[0091] 3. Third embodiment (particle sorting device)

[0092] Fig. 13 is a schematic conceptual diagram showing an example of the third embodiment. Fig. 14 is a schematic conceptual diagram showing another example of the third embodiment. A particle sorting device 30 according to this embodiment comprises a light irradiation unit 21, a light detection unit 22, a memory unit 11, a processing unit 12, and a sorting unit 31. It may also have other units such as a user interface 13 and a display unit 14, as necessary. In this embodiment, the memory unit 11, processing unit 12, user interface 13, display unit 14, light irradiation unit 21, and light detection unit 22 are the same as those described above, and therefore description thereof will be omitted here.

[0093] (1) Preparation section 31

[0094] The sorting unit 31 sorts particles based on the optical information detected by the light detection unit 22. More specifically, the sorting unit 31 sorts particles downstream of the flow path P based on the analysis results of particle size, shape, internal structure, etc., analyzed from light intensity data, for example.

[0095] The separation method will be explained in detail below for each figure.

[0096] In the particle sorting device 30 shown in Figure 13, droplets are generated from the outlet of the main flow path P13 by applying vibration to all or part of the main flow path P13 using, for example, a vibration element 31a that vibrates at a predetermined frequency. In this case, the vibration element 31a to be used is not particularly limited and can be selected as appropriate. One example is a piezoelectric vibration element. Furthermore, by adjusting the amount of liquid sent to the sample liquid flow path P11, the sheath liquid flow paths P12a and P12b, and the main flow path P13, the diameter of the outlet, the vibration frequency of the vibration element, etc., the size of the droplets can be adjusted, and droplets containing a constant amount of particles can be generated.

[0097] Next, the droplets are charged with a positive or negative electric charge (see reference numeral 31b in FIG. 13) based on the results of analysis of the particle size, shape, internal structure, etc., which are obtained based on the optical information detected by the photodetector 22. Then, the charged droplets are redirected to a desired direction by a counter electrode 31c to which a voltage is applied, and are then collected.

[0098] In addition, in the particle sorting device 30 shown in Figure 14, a sample liquid flow path P11 and sheath liquid flow paths P12a and P12b are formed on the substrate T, and three branch flow paths, a sorting flow path P14 and waste flow paths P15a and P15b, are provided downstream of the main flow path P13.Particles to be sorted that are determined to satisfy predetermined optical properties are taken into the sorting flow path P14, and particles not to be sorted that are determined not to satisfy the predetermined optical properties are allowed to flow into one of the two waste flow paths P15a and P15b without being taken into the sorting flow path P14, thereby allowing them to be sorted.

[0099] The particles to be sorted can be taken into the sorting channel P14 by any known method, for example, by generating a negative pressure in the sorting channel P14 using a vibration element 31a such as a piezoelectric element, and using this negative pressure to suck the sample liquid containing the particles to be sorted and the sheath liquid into the sorting channel P14. Although not shown, the particles to be sorted can also be taken into the sorting channel P14 by controlling or changing the direction of the laminar flow using an electromagnetic valve, a fluid stream (gas or liquid), or the like.

[0100] In this embodiment, in response to an instruction to exclude the flagged light intensity data, the sorting unit 31 sorts particles associated with the plurality of event data other than the event data including the flagged light intensity data, thereby enabling only particles having highly reliable data to be selected and sorted.

[0101] Specifically, for example, before sorting starts, the processing unit 12 performs processing to determine whether or not to perform sorting in the sorting unit 31 based on a setting as to whether or not to perform sorting on particles linked to the event data including the light intensity data to which the flag has been assigned. For example, if the setting is set to "not sort" particles linked to the event data including the light intensity data to which the flag has been assigned during sorting, it is possible to have a flow in which sorting is not performed even on events within the sorting target gate.

[0102] In this embodiment, the setting may be made in advance before sorting begins. Alternatively, the setting may be input by a user via the user interface 13. Furthermore, in this technology, the proportion of event data containing light intensity data to which the flag is assigned within all event data or the event data to be sorted can be displayed via the output processing unit 122. Based on the display result, the user can determine whether to exclude event data containing light intensity data to which the flag is assigned.

[0103] 4. Fourth embodiment (information processing method)

[0104] The information processing method according to this embodiment comprises a storage step and a processing step. Other steps may also be included as necessary. The method performed in the storage step is similar to the method performed in the storage unit 11 described above, and the method performed in the processing step is similar to the method performed in the processing unit 12 described above, so a description thereof will be omitted here.

[0105] The present technology can employ the following configuration. [1] a storage unit that stores event data consisting of light intensity data obtained by irradiating light onto one particle among the plurality of particles; a processing unit that processes a plurality of event data acquired from the plurality of particles; It consists of the storage unit stores a flag that is assigned when the light intensity data exceeds a threshold; The information processing device, wherein the processing unit performs processing on the plurality of event data other than the light intensity data to which the flag has been added, in response to an instruction to exclude the light intensity data to which the flag has been added. [2] The information processing device according to [1], wherein the flag is assigned when the light intensity exceeds an upper limit of detection in a photodetector that detects light from the particle and / or an upper limit of processing when processing the light intensity data. [3] The information processing device according to [2], wherein the flag is set when the input voltage range of analog-to-digital conversion is exceeded during analog-to-digital conversion of the signal from the photodetector. [4] The information processing device according to [2], wherein the flag is assigned when the upper limit value of data that can be held is exceeded when processing a digital signal. [5] The information processing device according to [2], wherein the flag is assigned when an upper limit value for data processing is exceeded when the light intensity data is processed. [6] The information processing device according to any one of [1] to [5], wherein the instruction is input by a user via a user interface. [7] the event data is composed of a plurality of light intensity data obtained by irradiating a single particle with a plurality of lights, The information processing device according to [1], wherein the processing unit performs processing on the plurality of event data other than the event data including the light intensity data in response to an instruction to exclude the light intensity data to which the flag is assigned. [8] The information processing device according to [7], wherein the processing unit outputs a ratio of event data that includes light intensity data to which the flag is assigned. [9] The information processing device according to [7] or [8], wherein the processing unit outputs a plot diagram created for the plurality of event data other than the event data including the light intensity data to which the flag has been assigned in response to an instruction to exclude the light intensity data to which the flag has been assigned.

[10] The information processing device according to [8], wherein the processing unit outputs a warning to a user when the ratio exceeds a threshold value.

[11] The information processing device according to any one of [7] to

[10] , wherein the processing unit performs a sorting process on particles associated with the plurality of event data other than the event data including the light intensity data in response to an instruction to exclude the light intensity data to which the flag is assigned.

[12] The information processing device according to [7], wherein the processing unit outputs a plot diagram created for the plurality of event data.

[13] The information processing device according to

[12] , wherein the processing unit calculates the ratio of event data including light intensity data to which the flag is assigned to a plurality of event data included in a gated area on the plot diagram.

[14] The information processing device according to

[13] , wherein the processing unit outputs a warning to a user when the ratio exceeds a threshold value.

[15] a light irradiation unit that irradiates light onto one particle of the plurality of particles; a light detection unit that detects light from the one particle; a storage unit that stores event data including light intensity data obtained from the light detection unit; a processing unit that processes a plurality of event data acquired from the plurality of particles; It consists of the storage unit stores a flag that is assigned when the light intensity data exceeds a threshold; the processing unit performs processing on the plurality of event data other than the light intensity data to which the flag is attached in response to an instruction to exclude the light intensity data to which the flag is attached. Particle analysis device.

[16] a light irradiation unit that irradiates light onto one particle of the plurality of particles; a light detection unit that detects light from the one particle; a storage unit that stores event data consisting of light intensity data obtained from the light detection unit; a processing unit that processes a plurality of event data acquired from the plurality of particles; It consists of the storage unit stores a flag that is assigned when the light intensity data exceeds a threshold; the processing unit processes the plurality of event data other than the light intensity data to which the flag is attached in response to an instruction to exclude the light intensity data to which the flag is attached; a fractionation unit that, in response to an instruction to exclude the flagged light intensity data, fractionates particles associated with the plurality of event data other than the event data including the flagged light intensity data; A particle sorting device comprising:

[17] a storage step of storing event data including light intensity data obtained by irradiating light onto one particle of the plurality of particles; a processing step of processing a plurality of event data acquired from the plurality of particles, In the storing step, a flag is stored that is assigned when the light intensity data exceeds a threshold value; In the processing step, processing is performed on the plurality of event data other than the light intensity data to which the flag has been assigned, in accordance with an instruction to exclude the light intensity data to which the flag has been assigned. [Explanation of symbols]

[0106] 10: Information processing device 11: Storage part 12: Processing section 121: arithmetic processing unit 122: Output processing unit 13: User Interface 14: Display section 20: Particle analysis device 21: Light irradiation unit 22: Light detection unit 30: Particle separation device 31: Preparative section

Claims

[Claim 1] a storage unit that stores event data consisting of light intensity data obtained by irradiating light onto one particle among the plurality of particles; a processing unit that processes a plurality of event data acquired from the plurality of particles; It consists of the storage unit stores a flag that is assigned when the light intensity data exceeds a threshold; The information processing device, wherein the processing unit performs processing on the plurality of event data other than the light intensity data to which the flag has been added, in response to an instruction to exclude the light intensity data to which the flag has been added.

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