Information processing device, particle analysis device, particle sorting device, and information processing method

Abstract

The present invention can enhance reliability of data analysis. For example, the present invention provides an information processing apparatus including a storage unit configured to store event data including light intensity data obtained by emitting light to a single particle among a plurality of particles, and a processing unit configured to process a plurality of event data items acquired from the plurality of particles. The storage unit stores a flag applied when the light intensity data exceeds a threshold value. In response to an instruction to omit the light intensity data to which the flag is applied, the processing unit performs processing on the plurality of event data items except for the light intensity data to which the flag is applied.

Description

Technical Field

[0001] This technology relates to information processing devices, particle analysis devices, particle sieving devices, and information processing methods. More specifically, this technology relates to information processing devices, particle analysis devices, particle sieving devices, and information processing methods for improving the reliability of data analysis. Background Technology

[0002] In the past, devices (e.g., flow cytometers) have been used that label particles such as cells with fluorescent dyes, illuminate the labeled particles with a laser beam, and detect the fluorescence or scattered light from the illuminated particles to measure various properties of the particles. In such devices, the light reaching the photodetector is converted into an electrical signal (voltage pulse) and digitized. The digital data is then subjected to statistical analysis and other methods under various parameters.

[0003] In recent years, multicolor measurements have been implemented, which involve labeling particles with multiple fluorescent dyes and using multiple photodetectors with different receiving wavelength bands to detect light emitted from different fluorescent dyes. In such multicolor measurements, each photodetector can receive fluorescence leaked from an unexpected fluorescent dye. To address this issue, fluorescence compensation is performed, in which the fluorescence intensity equivalent to the leaked fluorescence intensity is subtracted from the fluorescence intensity measured by each photodetector to improve the reliability of data analysis. Fluorescence compensation involves applying electrical or mathematical corrections to pulses on a dedicated circuit so that the fluorescence intensity measured by the photodetector becomes the true fluorescence intensity from the intended fluorescent dye.

[0004] For example, Patent Document 1 discloses a method in which the fluorescence intensity measured by each photodetector is represented as a vector, and the inverse of a predetermined leakage matrix is ​​applied to the vector to calculate the true fluorescence intensity of the target fluorescent dye.

[0005] Meanwhile, Patent Document 2 discloses a method in which the spectrum is approximated by the linear sum of simple staining spectra without resorting to the inverse of a predetermined leakage matrix in order to calculate the true fluorescence intensity from each fluorescent dye.

[0006] Existing technical documents

[0007] Patent documents

[0008] Patent Document 1: Japanese Patent Publication No. 2003-83894

[0009] Patent Document 2: Japanese Patent Publication No. 2011-232259 Summary of the Invention

[0010] The technical problem to be solved by the present invention

[0011] However, in existing devices, when the signal from the photodetector is converted from analog to digital, there may be photodetectors whose detection limit is exceeded by high signal levels. In this case, not only can the data of the signal input to that photodetector not be accurately obtained, but it also affects the signals acquired by other photodetectors through the fluorescence compensation process. This becomes a problem factor that reduces the overall reliability of the measurement data. Similarly, in this situation, gain adjustment is required for the purpose of measurement during the data measurement phase. Similarly, during the data analysis phase, gates are needed to exclude maximum values ​​from the measurement data plotted on each parameter axis.

[0012] Therefore, the main objective of this technology is to provide techniques for improving the reliability of data analysis.

[0013] Solution to the problem

[0014] According to the present technology, an information processing apparatus is provided, comprising: a storage unit configured to store event data including light intensity data obtained by irradiating one of a plurality of particles; and a processing unit configured to process a plurality of event data items obtained from the plurality of particles. The storage unit stores a flag assigned to the light intensity data when the light intensity data exceeds a threshold. Based on an instruction to exclude flagged light intensity data, the processing unit processes the plurality of event data items other than the flagged light intensity data.

[0015] According to the present technology, a particle analysis apparatus is also provided, comprising: a light irradiation unit configured to irradiate one of a plurality of particles with light; a light detection unit configured to detect light from the particles; a storage unit configured to store event data including light intensity data obtained from the light detection unit; and a processing unit configured to process a plurality of event data items obtained from the plurality of particles. The storage unit stores a flag assigned to the light intensity data when the light intensity data exceeds a threshold. Based on an instruction to exclude the flagged light intensity data, the processing unit processes the plurality of event data items other than the flagged light intensity data.

[0016] According to the present technology, a particle sieving apparatus is also provided, comprising: a light irradiation unit configured to irradiate one of a plurality of particles with light; a light detection unit configured to detect light from the particles; a storage unit configured to store event data including light intensity data obtained from the light detection unit; and a processing unit configured to process a plurality of event data items obtained from the plurality of particles. The storage unit stores a flag assigned to the light intensity data when the light intensity data exceeds a threshold. Based on an instruction to exclude the flagged light intensity data, the processing unit processes the plurality of event data items other than the flagged light intensity data. The apparatus also includes a sieving unit for sieving particles associated with the plurality of event data items other than the event data including light intensity data, based on an instruction to exclude the flagged light intensity data.

[0017] According to the present technology, an information processing method is also provided, comprising: a step of storing event data including light intensity data obtained by irradiating one of a plurality of particles; and a step of processing a plurality of event data items obtained from the plurality of particles. The storage step stores a flag assigned to the light intensity data if the light intensity data exceeds a threshold. Based on an instruction to exclude flagged light intensity data, the processing step processes the plurality of event data items other than the flagged light intensity data. Attached Figure Description

[0018] Figure 1 This is a schematic concept diagram depicting an example embodiment of an information processing apparatus according to the present technology.

[0019] Figure 2 In the diagram, subplot A is a plot depicting event data that does not include light intensity data exceeding a predetermined threshold, and subplot B is a plot depicting event data that includes light intensity data exceeding a predetermined threshold.

[0020] Figure 3 In the diagram, subplot A is a plot of event data that excludes light intensity data exceeding a predetermined threshold, and subplot B is a plot of event data that includes light intensity data exceeding a predetermined threshold.

[0021] Figure 4 In the diagram, subgraphs A, B, and C are examples of plotted diagrams.

[0022] Figure 5 This is a diagram depicting a display example shown by an output unit that outputs a ratio of event data including light intensity data set with the aforementioned markings.

[0023] Figure 6 This is a flowchart describing the processing example 1.

[0024] Figure 7 In the diagram, subgraphs A and B are schematic diagrams illustrating the data display examples of processing Example 1 and Example 2.

[0025] Figure 8 This is a flowchart describing the processing example 2.

[0026] Figure 9 This is a flowchart describing the processing of Example 3.

[0027] Figure 10 This is a flowchart describing the processing of Example 4.

[0028] Figure 11 This is a flowchart describing the processing of Example 5.

[0029] Figure 12This is a schematic conceptual diagram illustrating an example embodiment of a particle analysis apparatus according to the present technology.

[0030] Figure 13 This is a schematic conceptual diagram illustrating an example embodiment of a particle sieving apparatus according to the present technology.

[0031] Figure 14 This is a schematic concept diagram illustrating another embodiment of a particle sieving apparatus according to the present technology. Detailed Implementation

[0032] The following describes some preferred embodiments for implementing this technology.

[0033] It should be noted that the embodiments described below only illustrate how the technology can be implemented and should not be interpreted as limiting. The technology will be described in the following order.

[0034] 1. First Embodiment (Information Processing Device)

[0035] (1) Storage Unit 11

[0036] (2) Processing Unit 12

[0037] (3) User Interface 13

[0038] (4) Display Unit 14

[0039] (5) Example of processing performed by processing unit 12

[0040] <Processing Example 1>

[0041] <Processing Example 2>

[0042] <Processing Example 3>

[0043] <Processing Example 4>

[0044] <Processing Example 5>

[0045] 2. Second Embodiment (Particle Analysis Device)

[0046] (1) Light irradiation part 21

[0047] (2) Optical Detection Unit 22

[0048] (3) Processing Unit 12

[0049] 3. Third Embodiment (Particle Sieving Device)

[0050] (1) Screening section 31

[0051] 4. Fourth Implementation Method (Information Processing Method)

[0052] 1. First Embodiment (Information Processing Device)

[0053] Figure 1 This is a schematic conceptual diagram illustrating the first embodiment. The information processing apparatus 10 of this embodiment includes a storage unit 11 and a processing unit 12. The information processing apparatus 10 may also include other parts such as a user interface 13 and a display unit 14 as needed.

[0054] (1) Storage Unit 11

[0055] Storage unit 11 stores event data including light intensity data obtained by irradiating one of a plurality of particles. Storage unit 11 also stores a flag assigned to the light intensity data when the light intensity data exceeds a threshold.

[0056] In this technology, "particle" specifically refers to a microparticle, the type of which can be selected as needed. For this technology, microparticles can include biological particles, such as cells, cell clusters, microorganisms, and ribosomes, as well as synthetic particles, including gel particles, beads, latex particles, polymer particles, and industrial particles.

[0057] Bioparticles (also referred to as "biological particles") can include chromosomes, ribosomes, mitochondria, and organelles (cellular organelles) comprising different cells. Cells can include animal cells (e.g., blood cells) and plant cells. Specifically, cells can be blood cells or tissue cells. Blood cells can be floating cells, such as T cells and B cells. For example, tissue cells can be adherent cells cultured on a wall or adherent cells isolated from a tissue. Cell clusters can include, for example, spheroids and organoids. Microorganisms can include bacteria such as Escherichia coli, viruses such as tobacco mosaic virus, and fungi such as yeast. Bioparticles can also include biological macromolecules, such as nucleic acids, proteins, and their complexes. For example, these biological macromolecules can be extracted from cells or included in blood samples or other liquid samples.

[0058] Synthetic microparticles can be, for example, microparticles comprising organic or inorganic polymeric materials or metals. Organic polymeric materials can include polystyrene, styrene-divinylbenzene, and polymethyl methacrylate. Inorganic polymeric materials can include glass, silica, and magnetic materials. Metals can include gold colloids and aluminum. For example, these synthetic particles can be gel particles or beads. Specifically, these synthetic microparticles can be gel particles or beads formed from at least one or more combinations of oligonucleotides, peptides, proteins, and enzymes.

[0059] These particles can be spherical, substantially spherical, or non-spherical in shape. The size and mass of the particles can be selected as needed. For this technology, the particles can be provided with chemical or biological markers, such as fluorescent dyes or fluorescent proteins, as needed. Tags can be selected as needed. The tags can be conjugated to molecules that specifically react with the particles (e.g., antibodies, aptamers, DNA, or RNA).

[0060] For this technology, the particles are preferably biological particles, or more specifically, cells.

[0061] Fluorescent dyes used for labeling particles are not limited to any specific fluorescent dye. At least one known pigment used for staining biological products can be used. For example, available fluorescent dyes may include phycoerythrin (PE), fluorescein isothiocyanate (FITC), PE-Cy5, PE-Cy7, PE-Texas Red (registered trademark), allophycocyanin (APC), APC-Cy7, ethidium bromide, propidium iodide, Hoechst (registered trademark) 33258, Hoechst (registered trademark) 33342, DAPI (4',6-diamidinyl-2-phenylindole), acridine orange, chromomycin, styromycin, oligomycin, pyranin Y, thiazole orange, rhodamine 101, isothiocyanates, BCECF, BCECF-AM, C.SNARF-1 C.SNARF-1-AMA, jellyfish luminescent protein, Indo-1, Indo-1-AM, Fluo-3, Fluo-3-AM, Fura-2, Fura-2-AM, oxanol, TexasRed (registered trademark), Rhodamine 123, 10-N-nonanoic acid orange, fluorescein, fluorescein diacetate, carboxyfluorescein, carboxyfluorescein diacetate, carboxydichlorofluorescein, and carboxydichlorofluorescein diacetate. Derivatives of the fluorescent dyes listed above may also be used.

[0062] In this embodiment, light intensity data is generated by having a photodetector receive fluorescence or scattered light produced by irradiating the particle with light, and event data is generated based on the light intensity data. In the case where the particle is labeled with a fluorescent dye, the light intensity data is generated by having a photodetector receive fluorescence emitted from the fluorescent dye, which is excited by irradiating the particle with the fluorescent dye.

[0063] More specifically, when fluorescent or scattered light is received, the photodetector outputs an electrical signal corresponding to the received light. This output signal is then input to an analog-to-digital converter (ADC). The ADC is located downstream of the photodetector (on its output side) and connected to it. The electrical signal is an analog signal converted from the photoelectric signal by the photodetector. Each ADC converts each input electrical signal from analog to digital form. Then, each ADC outputs the digitized electrical signal to the downstream side.

[0064] The electrical signals output from each analog-to-digital converter (ADC) are input to a data detection circuit. The data detection circuit is located downstream (output side) of and connected to each ADC. The data detection circuit uses a specific one of the input electrical signals as a trigger signal for detecting particles. That is, when the value of the trigger signal meets a predetermined condition, the data detection circuit detects that each electrical signal has been detected from a particle. The data detection circuit reads the waveform of each input electrical signal and generates light intensity data by calculating the parameters (width, height, and area) of the read-in waveform. Furthermore, based on the values ​​of each parameter of the calculated waveform for each light intensity data item, the data detection circuit generates event data associated with a particle corresponding to that light intensity data. In this embodiment, the storage unit 11 stores the light intensity data and event data generated in the manner described above.

[0065] According to this technology, as described above, the storage unit 11 stores a flag assigned to the light intensity data when the light intensity data exceeds a threshold. Specifically, when the storage unit 11 receives a signal indicating that the light intensity data exceeds the threshold, it associates each light intensity data item with information indicating whether each light intensity data item exceeds the threshold. More specifically, the storage unit 11 assigns a flag to the light intensity data that exceeds the threshold and stores the flagged light intensity data.

[0066] For example, a flag can be assigned when the light from the particle detected by the aforementioned photodetector exceeds the detection limit of the photodetector. More specifically, for example, a flag can be assigned when the signal from the photodetector is converted from analog to digital by the aforementioned analog-to-digital converter circuit and exceeds the input voltage range of the analog-to-digital converter circuit, or when the digital signal is processed and exceeds the upper limit of the data retention capacity.

[0067] During digital signal processing, situations arise where the data holding capacity is exceeded, for example, by exceeding the upper limit of the data holding capacity (i.e., the upper limit that can be represented by bit width). In such cases, it is necessary to perform a process to remove (i.e., cut) the excess portion. This flag can be assigned during the cutting process.

[0068] The flag can also be assigned when processing light intensity data and the upper limit of data processing is exceeded. More specifically, the flag can be assigned when the upper limit of the capacity for holding light intensity data is exceeded when generating event data based on light intensity data, or when the upper limit of the capacity for holding data for conversion to the data transmission format is exceeded when light intensity data or event data is transmitted from the information processing device to the data analysis device (e.g., a personal computer or server).

[0069] In this embodiment, the event data may include multiple light intensity data items obtained by irradiating the particle with multiple light beams. In this case, multiple light beams may be emitted from multiple light sources capable of irradiating excitation beams with different wavelengths. In this embodiment, for example, light beams from multiple light sources may be directed to different locations, such that the light from the particle may be detected by different photodetectors to provide multiple light intensity data items.

[0070] (2) Processing Unit 12

[0071] The processing unit 12 processes multiple event data acquired from multiple particles. Furthermore, based on an instruction to remove the marked light intensity data, the processing unit 12 processes multiple event data items preceding the marked light intensity data.

[0072] When performing analog-to-digital conversion on signals from a photodetector, existing devices lack a means to exclude light intensity data exceeding the aforementioned threshold. Figure 2 In the diagram, subplot A depicts event data that excludes light intensity data exceeding a threshold, and subplot B depicts event data that includes light intensity data exceeding a threshold. Therefore, the existing device cannot exclude light intensity data exceeding the threshold. Consequently, there is no alternative other than adjusting the gain used for measurement during the data measurement phase in a manner that does not exceed the threshold.

[0073] exist Figure 3 In the diagram, subplot A is a plot showing the case where event data exceeding a threshold is excluded, and subplot B is a plot showing the case where event data exceeding a threshold is included. For example, in the case of event data exceeding a threshold during the data analysis phase, existing devices can only allow the mixing of light intensity data that exceeds the threshold more or less by creating a gate to exclude the maximum value data along each parameter axis of the plotted measurement data.

[0074] Furthermore, if a photodetector detects light intensity data exceeding a threshold, the signal data input to that photodetector cannot be accurately obtained. Additionally, signals acquired from other photodetectors during fluorescence compensation processing are also affected. Therefore, the reliability of data analysis suffers.

[0075] Conversely, according to this technology, the processing unit 12 processes multiple event data items other than the marked light intensity data according to an instruction to exclude marked light intensity data. This technology therefore provides a means of excluding marked light intensity data when displaying, analyzing, or processing acquired event data. This technology allows data that may reduce the reliability of results to be excluded from measurement data as needed, thereby improving the reliability of data analysis.

[0076] More specifically, when creating information for performing fluorescence compensation such as separation or compensation (e.g., spectral reference, compensation matrix, etc.), it is determined whether to exclude the labeled light intensity data. The data generated based on the determination result is used to obtain data with higher reliability. Note that this process will be discussed later in "(5) Example of processing performed by processing unit 12".

[0077] Using this embodiment, when the event data includes multiple light intensity data items obtained by irradiating multiple light beams onto particles, the processing unit 12 can process multiple event data items other than the event data including the marked light intensity data according to the instruction to exclude the marked light intensity data.

[0078] 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 ratio of event data including the marked light intensity data. The output processing unit 122 also performs processing to output the ratio of invention data including the marked light intensity data and / or output drawing processing.

[0079] For example, the arithmetic processing unit 121 in this embodiment can calculate the ratio of event data including marked light intensity data to event data (all event data) including a series of multiple light intensity data items obtained by irradiating each of multiple particles with multiple beams. Alternatively, the arithmetic processing unit 121 can calculate the ratio of event data including marked light intensity data to multiple event data items included in a selected area on a drawing output by the output processing unit 122, which will be discussed later. When calculated, these ratios provide indicators by which the user can determine whether to exclude the marked light intensity data from all event data or from multiple event data items included in a selected area on the drawing.

[0080] In this embodiment, the output processing unit 122 may also output, for example, a ratio of event data including labeled light intensity data from multiple event data items obtained from multiple particles used for analysis. This ratio output provides an indicator, through which the user can determine whether to exclude, for example, labeled light intensity data.

[0081] Furthermore, the output processing unit 122 can output the ratio of event data, including the marked light intensity data, to the display unit 14, which will be discussed later. Figure 5 This is a diagram illustrating a display example where a display unit 14 displays a ratio of event data including the marked light intensity data. In this case, by referring to the displayed ratio, the user can input via the user interface 13 to determine whether to exclude the marked light intensity data.

[0082] Furthermore, if the ratio of event data including the labeled light intensity data exceeds a threshold, the output processing unit 122 can output a warning to the user. Using this technique, the threshold can be set by the user as needed. The warning can be displayed on the display unit 14 (discussed later) or issued audibly to alert the user.

[0083] In this embodiment, the output processing unit 122 can also output a graph created for multiple event data items obtained from multiple particles used for analysis. In this case, the arithmetic processing unit 121 also calculates the ratio of the event data including the marked light intensity data to the multiple event data items included in the gated area on the graph.

[0084] Here, the ratio of event data, including the marked light intensity data, can be output by the output processing unit 122 in a state displayed on the plot, or the ratio of only the event data can be output on a different screen. For display on the plot, the ratio of the event data relative to multiple event data items included in the gating area can be displayed on a newly created sub-plot. This output allows for an assessment of the reliability of event data that the user wishes to analyze in detail. This enables more precise analysis.

[0085] Furthermore, in this embodiment, the output processing unit 122 may issue a warning to the user if the ratio of the event data including the marked light intensity data to a plurality of event data items included in a gating area on the drawing exceeds a threshold. Using this technique, the threshold can be set by the user as needed. The warning may be displayed on the display unit 14, discussed later, or may be audibly issued to alert the user.

[0086] In this embodiment, based on the instruction to exclude labeled light intensity data, the processing unit 12 can perform a sieving process on particles related to multiple event data items other than event data including labeled light intensity data. This method allows for the selective sieving of only particles with high-reliability data. It should be noted that this process will be discussed later in "(1) Sieving Unit 31".

[0087] Furthermore, the processing unit 12 in this embodiment can perform processing to switch between methods for displaying spectral graphs. A spectral graph is a graph displaying light intensity data obtained by irradiating a single beam of light in different wavelength bands of a photodetector. For example, when displaying data based on the frequency of detected event data including predetermined light intensity data, the data can be color-coded. In the case of irradiating particles with multiple beams, a band diagram can be displayed, wherein spectral graphs corresponding to different beams are arranged (see [link to relevant documentation]). Figure 4 More specifically, the process of switching between display methods includes switching from a method that displays all light intensity data obtained through analysis in a color-coded manner according to their frequencies to a method that does not display data whose frequencies do not exceed a given threshold.

[0088] exist Figure 4 In the diagram, subplot A depicts a plot showing all event data in a color-coded manner according to their frequencies; subplot B depicts a plot that does not display data whose frequencies do not exceed a predetermined threshold; and subplot C depicts a plot that does not display data whose frequencies do not exceed a threshold set above the threshold in subplot B. When the display method is switched in this way, it is easier to see the fluorescence spectral waveforms of the main population (e.g., a single or multiple cell populations).

[0089] Furthermore, in this embodiment, the threshold can be changed according to the user's expectations. Alternatively, a fixed threshold can be preset. If the user needs to change the threshold, they can enter a new threshold or select one of the available thresholds using a slider.

[0090] (3) User Interface 13

[0091] User interface 13 allows user input. Through user interface 13, the user can access and control components of the information processing apparatus of this embodiment. Note that user interface 13 is not mandatory for this technology and can be replaced by an externally connected operating device. For example, a mouse and keyboard can be used as user interface 13.

[0092] In this embodiment, the user can input and execute commands via the user interface 13 to exclude marked light intensity data. Commands to exclude marked light intensity data can be executed via the user interface 13 as needed, not only when the output processing unit 122 outputs a ratio of event data including the marked light intensity data or outputs a warning based on that ratio. For example, commands can be executed appropriately by selecting a button displayed on the screen.

[0093] (4) Display Unit 14

[0094] Display unit 14 can display, for example, information output from processing unit 12, as well as all analysis-related matters generated by or output from any component of the device. Note that display unit 14 is not mandatory for this technology and can be replaced by an externally connected display device. For example, a display unit and a printer can be used as display unit 14.

[0095] (5) Example of processing performed by processing unit 12

[0096] The following describes some examples of the processing performed by the processing unit 12 with reference to the attached flowchart.

[0097] <Processing Example 1>

[0098] Figure 6 This is a flowchart describing Processing Example 1. Processing Example 1 is a process for registration as a spectral reference. The spectral reference is formed by a simple staining spectrum, which is used, for example, in a separation process. The simple staining spectrum is the fluorescence wavelength distribution of a fluorescent dye, i.e., light intensity data obtained when a photodetector receives fluorescence emitted from the fluorescent dye, each fluorescent dye labeling a particle excited by light.

[0099] Unmixing is a fluorescence compensation method in which particles labeled with multiple fluorescent dyes are measured in multiple colors to provide a measurement spectrum. This spectrum is approximated by a linear sum of the simple staining spectra using a weighted least squares (WLSM) method to obtain the true light intensity data derived from each fluorescent dye. Performing the unmixing process separates the spectral information about overlapping fluorescent dyes into information about individual fluorescent dyes. Fluorescent reagents and fluorescent proteins whose fluorescence wavelength peaks are very similar to each other can then be separated and analyzed with high precision and in a highly reproducible manner.

[0100] First, the processing unit 12 receives the marked light intensity data (S1). Note that the light intensity data in this case is formed by the simple staining spectrum described above. The arithmetic processing unit 121 then calculates the ratio of the event data including the marked light intensity data to the event data (all event data) including, for example, a series of multiple light intensity data items obtained by continuously irradiating each of multiple particles with multiple beams. The output processing unit 122 outputs the calculated ratio (S2). At this time, if the ratio exceeds a threshold (S3), the output processing unit 122 outputs a warning to the user (S4). Then it determines whether to exclude the marked light intensity data (S5). Note that this determination can be made by the user. In this case, the user inputs a command via the user interface 13.

[0101] If it is determined that the marked light intensity data will be excluded (S5), the processing unit 12 excludes the marked light intensity data (S5) and provides data display (S7). The data display method is not limited to any particular method; for example, the data can be displayed in any of the following on the display unit 14: plots (including one-dimensional, two-dimensional, and three-dimensional plots), spectra, and histograms (including one-parameter histograms, two-parameter histograms (cell plots, dot plots), and three-parameter histograms). Specifically, the output processing unit 122 outputs plots created for multiple event data items other than event data including the marked light intensity data. Figure 7 Subplots A and B are schematic diagrams illustrating data display examples of Processing Example 1 and Processing Example 2 discussed later. Subplot A is a plot where the vertical axis represents light intensity and the horizontal axis represents the photodetector channel. Subplot B is a histogram where the vertical axis represents the number of events and the horizontal axis represents the light intensity at the fluorescence wavelength peak of the labeled particle. On the other hand, if it is determined that the light intensity data of the labeled particle is not excluded (S5), the processing unit 12 provides data display without excluding the light intensity data of the labeled particle (S7).

[0102] In the displayed data, both the positive and negative populations are selected (S8). Note that the selection process can be performed by the user. In this case, the user inputs instructions via the user interface 13. Then, the processing unit 12 obtains the average light intensity value of each of the positive and negative populations and performs differential processing on them (S9). Finally, the data obtained from the differential processing is registered in the storage unit 11 as a spectral reference (S10).

[0103] <Processing Example 2>

[0104] Figure 8 This is a flowchart describing Processing Example 2. Similar to Processing Example 1, Processing Example 2 is a process registered as a spectral reference.

[0105] First, the processing unit 12 receives the marked light intensity data (S11). Note that the light intensity data in this case is formed from the simple stained spectrum described above. Then, the processing unit 12 provides data display (S12). The data display method is as described above. In this case, for example, the output processing unit 122 can output a plot created for multiple event data items. In the displayed data, both the positive and negative groups are gated (S13). Note that the gating can be performed by the user. In this case, the user inputs instructions via the user interface 13.

[0106] The arithmetic processing unit 121 then calculates the ratio of the event data, including the marked light intensity data, to the ratio of the multiple event data items included in the gating area on the graph output by the output processing unit 122. The output processing unit 122 outputs the calculated ratio (S14). At this time, if the ratio exceeds a threshold (S15), the output processing unit 122 outputs a warning to the user (S16). Then it determines whether to exclude the marked light intensity data (S17). Note that this determination can be made by the user. In this case, the user inputs a command via the user interface 13.

[0107] If it is determined that the labeled light intensity data should be excluded (S17), the labeled light intensity data is excluded (S18). Then, the processing unit 12 obtains the average light intensity value of each of the positive and negative groups and performs differential processing on it (S19). Finally, the data obtained from the differential processing is registered in the storage unit 11 as a spectral reference (S20). On the other hand, if it is determined that the labeled light intensity data should not be excluded (S17), the processing unit 12 performs the above differential processing without excluding the labeled light intensity data (S19), and registers the resulting data in the storage unit 11 as a spectral reference (S20).

[0108] <Processing Example 3>

[0109] Figure 9 This is a flowchart describing Processing Example 3. Processing Example 3 is a process where compensation is performed automatically. The compensation here involves compensating for leaked fluorescence. In the case of two-color measurements, for example, where two fluorescence wavelengths overlap, the compensation method involves electrically or mathematically compensating for the amount of mutual leakage. Methods for mathematically performing fluorescence compensation may include, for example, representing the fluorescence intensity measured by each photodetector as a vector and applying the inverse of a predetermined leakage matrix to that vector to calculate the true fluorescence intensity of the desired fluorescent dye. The leakage matrix generated by analyzing the simple staining spectrum described above is a column vector array formed by the fluorescence wavelength distribution of the fluorescent dyes involved.

[0110] First, the processing unit 12 receives the marked light intensity data (S21). Note that the light intensity data in this case includes the simple staining spectrum described above. The arithmetic processing unit 121 then calculates the ratio of the event data including the marked light intensity data to the event data (all event data) formed by a series of multiple light intensity data items obtained by irradiating multiple beams of light to multiple particles in succession. The output processing unit 122 outputs the calculated ratio (S22). At this time, if the ratio exceeds a threshold (S23), the output processing unit 122 outputs a warning to the user (S24). Then, it is determined whether to exclude the marked light intensity data (S25). Note that this determination can be made by the user. In this case, the user inputs a command via the user interface 13.

[0111] If it is determined that the marked light intensity data should be excluded (S25), the marked light intensity data is excluded (S26). The processing unit 12 then provides data display according to the instruction to exclude the marked light intensity data (S27). The data display method is not limited to any particular method; for example, the data can be displayed in any of the following on the display unit 14: plots (including one-dimensional, two-dimensional, and three-dimensional plots), spectra, and histograms (including one-parameter histograms, two-parameter histograms (cell plots, dot plots), and three-parameter histograms). Similarly, in this case, for example, the output processing unit 122 can output plots created for multiple event data items (excluding event data that includes the marked light intensity data). On the other hand, if it is determined that the marked light intensity data is not excluded (S25), the processing unit 12 provides the above-mentioned data display without excluding the marked light intensity data (S27).

[0112] In the displayed data, both the positive and negative groups are gated (S28). Note that the gateding process can be performed by the user. In this case, the user inputs instructions via user interface 13. Finally, matrix calculations are performed using the data obtained as a result of the gateding process (S29).

[0113] <Processing Example 4>

[0114] Figure 10 This is a flowchart describing processing example 4. Similar to processing example 3, processing example 4 is the processing under the condition of automatic compensation.

[0115] First, the processing unit 12 receives the labeled light intensity data (S31). Note that the light intensity data in this case includes the simple staining spectrum described above. Then, the processing unit 12 provides data display (S32). The data display method is as described above. In this case, for example, the output processing unit 122 can output a graph created for multiple event data items. Then, in the displayed data, the positive and negative groups are gated respectively (S33). Note that the gatening process can be performed by the user. In this case, the user inputs instructions via the user interface 13.

[0116] The arithmetic processing unit 121 then calculates the ratio of the event data including the marked light intensity data to the multiple event data items included in the gating area on the graph output by the output processing unit 122. The output processing unit 122 outputs the calculated ratio (S34). If this ratio exceeds a threshold (S35), the output processing unit 122 outputs a warning to the user (S36). Then it determines whether to exclude the marked light intensity data (S37). Note that this determination can be made by the user. In this case, the user inputs a command via the user interface 13.

[0117] If it is determined that the light intensity data of the marker will be excluded (S37), the light intensity data of the marker is excluded (S38). Finally, the obtained data is used to perform matrix calculation (S39). On the other hand, if it is determined that the light intensity data of the marker is not excluded (S37), the processing unit 12 performs matrix calculation using the obtained data without excluding the light intensity data of the marker (S39).

[0118] <Processing Example 5>

[0119] Figure 11 This is a flowchart describing processing example 5. Processing example 5 describes the process in the case where the user manually performs compensation.

[0120] First, the processing unit 12 receives the labeled light intensity data (S41). Note that the light intensity data in this case includes the measured spectrum. The measured spectrum refers to the light intensity data obtained by causing a photodetector to receive fluorescence emitted from multiple fluorescent dyes, which are excited by irradiating light with particles multipletically labeled with these fluorescent dyes whose fluorescence wavelengths overlap. The arithmetic processing unit 121 then calculates the ratio of the event data including the labeled light intensity data to the event data (all event data) formed by a series of multiple light intensity data items obtained by continuous measurement of multiple particles. The output processing unit 122 outputs the calculated ratio (S42). At this time, if the ratio exceeds a threshold (S43), the output processing unit 122 outputs a warning to the user (S44). Then it determines whether to exclude the labeled light intensity data (S45). Note that this determination can be made by the user. In this case, the user inputs a command via the user interface 13.

[0121] If it is determined that the marked light intensity data will be excluded (S45), the marked light intensity data is excluded (S46). The processing unit 12 then provides data display according to the instruction to exclude the marked light intensity data (S47). The data display method is not limited to any particular method; for example, the data can be displayed in any of the following on the display unit 14: plots (including one-dimensional, two-dimensional, and three-dimensional plots), spectra, and histograms (including one-parameter histograms, two-parameter histograms (cell plots, dot plots), and three-parameter histograms). In the same case, the output processing unit 122 can output a plot created for multiple event data items (excluding event data that includes the marked light intensity data). On the other hand, if it is determined that the marked light intensity data will not be excluded (S45), the processing unit 12 provides the above-mentioned data display without excluding the marked light intensity data (S47). Finally, the user performs visual correction processing on the displayed data (S48).

[0122] 2. Second Embodiment (Particle Analysis Device)

[0123] Figure 12 This is a schematic conceptual diagram illustrating the second embodiment. The particle analysis apparatus 20 of this embodiment includes a light irradiation unit 21, a light detection unit 22, a storage unit 11, and a processing unit 12. The particle analysis apparatus 20 may further include other components, such as a user interface 13 and a display unit 14, as needed. In this embodiment, the storage unit 11, user interface 13, and display unit 14 are similar to those described above, and therefore will not be discussed further here.

[0124] (1) Light irradiation part 21

[0125] The light irradiation unit 21 illuminates one of the plurality of particles with light (e.g., excitation light). The light irradiation unit 21 may include a light source emitting the excitation light and an objective lens focusing the excitation light onto the particle. Those skilled in the art can appropriately select a suitable light source. For example, the light source may be a laser diode, an SHG laser, a solid-state laser, a gas laser, a high-brightness LED, or a combination of two or more of these devices. In addition to the light source and objective lens, the light irradiation unit 21 may also include other optical elements. The light irradiation unit 21 may, for example, illuminate a single location within an optical detection region, or illuminate each of a plurality of locations therein.

[0126] In this embodiment, the light irradiation unit 21 may have multiple light sources as described above, so as to irradiate excitation beams with different wavelengths.

[0127] (2) Optical Detection Unit 22

[0128] The light detection unit 22 detects light from a single particle. More specifically, the light detection unit 22 detects scattered light and / or fluorescence emitted from a particle irradiated by the light irradiation unit 21. The light detection unit 21 may include, for example, a photodetector and a condenser lens that focus the fluorescence and / or scattered light emitted from the particle. The photodetector may be a PMT, a photodiode, a CCD, or a CMOS, but is not limited thereto. Depending on the need, the light detection unit 22 may include other optical elements besides the condenser lens and the detector. For example, the light detection unit 22 may further include a spectral section. For example, the optical portion including the beam splitter may be a grating, a prism, and a filter. The beam splitter is capable of separating and detecting light of the wavelength to be detected from beams of other wavelengths.

[0129] The fluorescence detected by the light detection unit 22 can be fluorescence emitted from the particles themselves or from a material labeled with the particles (e.g., a fluorescent material), but is not limited thereto. The scattered light detected by the light detection unit 22 can be forward scattered light, side scattered light, Rayleigh scattering, Mie scattering, or a combination of these beams.

[0130] (3) Processing Unit 12

[0131] In this embodiment, in addition to the processes described above, the processing unit 12 may also perform the following processes.

[0132] The processing unit 12, connected to the light detection unit 22, can analyze the optical information detected therefrom. More specifically, for example, given optical information related to the light received from the light detection unit 22 (e.g., detection values ​​of fluorescence or scattered light), the processing unit 12 calculates characteristic quantities representing the size, shape, and internal structure of each particle.

[0133] It should be noted that, using this technology, external analysis devices can be used for analysis. Specifically, a personal computer or CPU can be used to perform the analysis, store the results as a program in hardware resources equipped with recording media (e.g., non-volatile memory (USB memory), HDD, CD), etc.), and allow the program to run. Furthermore, external analysis devices can be connected to components of the device via a network.

[0134] 3. Third Embodiment (Particle Sieving Device)

[0135] Figure 13 This is a schematic conceptual diagram illustrating an example of a third embodiment. Furthermore, Figure 14 This is a schematic conceptual diagram illustrating another example of the third embodiment. The particle sieving apparatus 30 of this embodiment includes a light irradiation unit 21, a light detection unit 22, a storage unit 11, a processing unit 12, and a sieving unit 31. The particle sieving apparatus 30 may further include other components, such as a user interface 13 and a display unit 14, as needed. In this embodiment, the storage unit 11, processing unit 12, user interface 13, display unit 14, light irradiation unit 21, and light detection unit 22 are similar to those described above, and therefore will not be discussed further.

[0136] (1) Screening section 31

[0137] The sieving unit 31 sieves particles based on optical information detected by the light detection unit 22. More specifically, for example, the sieving unit 31 sieves particles downstream of the flow channel P from light intensity data based on analysis results including particle size, shape, and internal structure.

[0138] The screening and separation method is described in detail below with reference to the separate accompanying drawings.

[0139] Figure 13 The particle sieving device 30 in the process vibrates all or part of the main flow path P13 by using a vibrating element 31a that vibrates at a redefined vibration frequency, thereby discharging droplets from the outlet of the main flow path P13. Note that the vibrating element 31a used in this case is not limited to any particular type and can be selected as needed. For example, a piezoelectric vibrating element can be used. Furthermore, by adjusting the amount of liquid supplied to the sample liquid flow path P11, the sheath liquid flow paths P12a and P12b, and the main flow path P13, and by adjusting the diameter of the outlet and the vibration frequency of the vibrating element, droplets with a fixed number of particles can be generated by adjusting the droplet size.

[0140] Then, based on the optical information detected by the photodetector 22, and based on the analysis results such as the particle size, shape, and internal structure, the droplet is determined to be positively or negatively charged (see...). Figure 13(The content is indicated by reference symbol 31b). The charged droplet is separated by applying a voltage to the opposite electrode 31c, which changes its path in the desired direction.

[0141] In addition, Figure 14 In the particle sieving apparatus 30, a sample liquid flow path P1 and sheath liquid flow paths P12a and P12b are formed on a substrate T. Three branch flow paths, including a sieving flow path P14 and disposal flow paths P15a and P15b, are arranged downstream of the main flow path P13. Particles determined to meet predetermined optical characteristics and targeted for sieving are carried into the sieving flow path P14. Particles determined not to meet the predetermined optical characteristics are not carried into the sieving separation flow path P14 and are guided to either of the two disposal flow paths P15a and P15b. This is how sieving is performed.

[0142] Any known method can be used to draw the target particles for sieving into the sieving channel P14. Alternatively, a vibrating element 31a, such as a piezoelectric element, can be used to generate a negative pressure inside the sieving channel P14, which will draw the sample liquid and sheath liquid containing the target particles into the sieving flow channel P14. As another alternative, valve electromagnetic force, fluid flow (gas or liquid), etc., can be used to control or change the laminar flow direction, thereby drawing the target particles for sieving into the sieving flow channel P14.

[0143] In this embodiment, the screening unit 31 screens particles related to multiple event data items other than event data including light intensity data, according to an instruction to exclude marked light intensity data. This allows for the selective screening and separation of only particles with high-reliability data.

[0144] Specifically, before sorting begins, the processing unit 12 can determine whether to perform a screening process based on a setting that allows the screening unit 31 to screen particles associated with event data including marked light intensity data. For example, if there is a setting during sorting that prevents the screening of particles associated with event data including marked light intensity data, the flow can be arranged such that events within the sorting gate will not be sorted.

[0145] In this embodiment, settings can be made before sorting begins. This setting can alternatively be performed by the user via user interface 13. Similarly, for this technology, the ratio of event data including marked light intensity data to all event data or to event data categorized can be displayed via output processing unit 122. Based on the displayed results, the user can determine whether to exclude event data including marked light intensity data.

[0146] 4. Fourth Implementation Method (Information Processing Method)

[0147] The information processing method of this embodiment includes a storage step and a processing step. Other steps may be included in this method as needed. The method performed in the storage step is the same as the method performed in the storage unit 11, and the method performed in the processing step is the same as the method performed in the processing unit 12. Therefore, these methods will not be discussed further.

[0148] It should be noted that this technology can preferably be implemented in the following configuration. (1)

[0150] An information processing apparatus, comprising:

[0151] The storage unit is configured to store event data, including light intensity data obtained by irradiating one of a plurality of particles; and

[0152] The processing unit is configured to process multiple event data items acquired from the plurality of particles, wherein,

[0153] The storage unit stores the flags assigned to the light intensity data when the light intensity data exceeds a threshold, and...

[0154] According to the instruction to exclude the light intensity data of the marker, the processing unit processes the plurality of event data items other than the light intensity data of the marker. (2)

[0156] According to the information processing apparatus of (1), the marker is provided when the detection limit of the photodetector that detects light from the particle is exceeded and / or the processing limit of the ability to process the light intensity data is exceeded. (3)

[0158] According to the information processing apparatus of (2), when the signal from the photodetector is converted from analog to digital, a mark is assigned if the input voltage range of the analog-to-digital conversion is exceeded. (4)

[0160] According to the information processing apparatus of (2), a mark is assigned when the capacity for holding data exceeds the upper limit during the processing of digital signals. (5)

[0162] According to the information processing apparatus of (2), a mark is assigned when the processing capacity for light intensity data exceeds the upper limit of the data processing capability. (6)

[0164] The information processing apparatus according to any one of (1) to (5), wherein the instructions are input by the user via a user interface. (7)

[0166] According to the information processing apparatus described in (1), wherein

[0167] The event data includes multiple light intensity data items obtained by irradiating the particle with multiple beams of light, and,

[0168] According to the instruction to exclude the marked light intensity data, the processing unit processes multiple event data items other than event data including light intensity data. (8)

[0170] According to the information processing device of (7), the processing unit outputs a ratio of event data including marked light intensity data. (9)

[0172] According to the information processing apparatus of (7) or (8), the processing unit outputs a drawing created for multiple event data items other than event data including the light intensity data of the marker, according to the instruction to exclude the light intensity data of the marker. (10)

[0174] According to the information processing apparatus described in (8), if the ratio exceeds a threshold, the processing unit outputs a warning to the user. (11)

[0176] According to any one of (7) to (10) the information processing apparatus, wherein, according to the instruction to exclude the light intensity data of the marker, the processing unit performs sieving processing on particles related to a plurality of event data items other than event data including light intensity data. (12)

[0178] According to the information processing apparatus of (7), the processing unit outputs a drawing created for multiple event data items. (13)

[0180] According to the information processing apparatus of (12), the processing unit calculates the ratio of event data including marked light intensity data to multiple event data items included in a gating area on a drawing. (14)

[0182] According to the information processing apparatus described in (13), if the ratio exceeds a threshold, the processing unit outputs a warning to the user. (15)

[0184] A particle analysis device, comprising:

[0185] The light irradiation section is configured to irradiate one of a plurality of particles;

[0186] A photodetector is configured to detect light from the particles;

[0187] The storage unit is configured to store event data, including light intensity data obtained from the light detection unit; and

[0188] The processing unit is configured to process multiple event data items acquired from the plurality of particles, wherein,

[0189] The storage unit stores the flags assigned to the light intensity data when the light intensity data exceeds a threshold, and

[0190] According to the instruction to exclude the marked light intensity data, the processing unit processes the plurality of event data items other than the marked light intensity data. (16)

[0192] A particle sieving device, comprising:

[0193] The light irradiation section is configured to irradiate one of a plurality of particles;

[0194] A photodetector is configured to detect light from the particles;

[0195] The storage unit is configured to store event data, including light intensity data obtained from the light detection unit; and

[0196] The processing unit is configured to process multiple event data items acquired from the plurality of particles, wherein,

[0197] The storage unit stores the flags assigned to the light intensity data when the light intensity data exceeds a threshold.

[0198] According to the instruction to exclude the light intensity data of the marker, the processing unit processes the plurality of event data items other than the light intensity data of the marker, and,

[0199] The sieving section is further configured to sieve particles associated with multiple event data items other than event data including light intensity data, according to instructions to exclude marked light intensity data. (17)

[0201] An information processing method, comprising:

[0202] The steps of storing event data include obtaining light intensity data by illuminating one of a plurality of particles; and

[0203] The step of processing multiple event data items obtained from the plurality of particles, wherein,

[0204] The storage step stores a flag assigned to the light intensity data when the light intensity data exceeds a threshold, and...

[0205] According to the instruction to exclude the marked light intensity data, the processing step processes the plurality of event data items other than the marked light intensity data.

[0206] Reference number list

[0207] 10 Information Processing Devices

[0208] 11 Storage Department

[0209] 12 Processing Department

[0210] 121 Arithmetic Processing Department

[0211] 122 Output Processing Unit

[0212] 13 User Interface

[0213] 14 Display Section

[0214] 20-Particle Analysis Device

[0215] 21Light irradiation part

[0216] 22 Optical Detection Department

[0217] 30-particle screening device

[0218] 31 Screening section.

Claims

1. An information processing apparatus comprising: a storage configured to store event data including light intensity data obtained by irradiating light to one of a plurality of particles; and a processing section configured to process a plurality of event data items acquired from the plurality of particles, wherein the storage stores a flag given to the light intensity data in a case where the light intensity data exceeds a threshold value, and the processing section processes a plurality of event data items other than the light intensity data to which the flag is given, according to an instruction to exclude the light intensity data to which the flag is given.

2. The information processing apparatus according to claim 1, wherein The flag is given in a case where a detection upper limit of a light detector that detects light from the particle is exceeded and / or a processing upper limit of a processing data is exceeded, when the light intensity data is processed.

3. The information processing apparatus according to claim 2, wherein The flag is given in a case where a range of an input voltage of an analog-digital conversion of a signal from the light detector is exceeded, when the signal from the light detector is analog-digital converted.

4. The information processing apparatus according to claim 2, wherein The flag is given in a case where an upper limit of a capacity of holding data is exceeded, when a digital signal is processed.

5. The information processing apparatus according to claim 2, wherein The flag is given in a case where an upper limit of a capacity of the processing data is exceeded, when the light intensity data is processed.

6. The information processing apparatus according to claim 1, wherein The instruction is input by a user via a user interface. 7.The information processing apparatus according to claim 1, wherein the event data includes a plurality of light intensity data items obtained by irradiating a plurality of light beams to the particle, and the processing section processes a plurality of event data items other than event data including the light intensity data to which the flag is given, according to an instruction to exclude the light intensity data to which the flag is given.

8. The information processing apparatus according to claim 7, wherein The processing section outputs a ratio of event data including the light intensity data to which the flag is given.

9. The information processing apparatus according to claim 7, wherein The processing section outputs a plot created for a plurality of event data items other than event data including the light intensity data to which the flag is given, according to the instruction to exclude the light intensity data to which the flag is given.

10. The information processing apparatus according to claim 8, wherein The processing section outputs a warning to a user in a case where the ratio exceeds a threshold value.

11. The information processing apparatus according to claim 7, wherein The processing section performs a screening process on particles associated with a plurality of event data items other than event data including the light intensity data to which the flag is given, according to the instruction to exclude the light intensity data to which the flag is given.

12. The information processing apparatus according to claim 7, wherein The processing section outputs a plot created for the plurality of event data items.

13. The information processing apparatus according to claim 12, wherein The processing section calculates a ratio of the event data including the light intensity data to which the flag is given with respect to a plurality of event data items included in a strobe region on the plot.

14. The information processing apparatus according to claim 13, wherein The processing section outputs a warning to a user in a case where the ratio exceeds a threshold value. 15.An apparatus for particle analysis comprising: a light irradiation section configured to irradiate light to one of a plurality of particles; a light detection section configured to detect light from the particle; a storage configured to store event data including light intensity data obtained from the light detection section; and a processing section configured to process a plurality of event data items acquired from the plurality of particles, wherein the storage stores a flag given to the light intensity data in a case where the light intensity data exceeds a threshold value, and the processing section processes a plurality of event data items other than the light intensity data to which the flag is given, according to an instruction to exclude the light intensity data to which the flag is given. 16.An apparatus for particle screening comprising: a light irradiation section configured to irradiate one of a plurality of particles with light; a light detection section configured to detect light from the particle; a storage section configured to store event data including light intensity data obtained from the light detection section; and a processing section configured to process a plurality of event data items acquired from the plurality of particles, wherein the storage section stores a mark assigned to the light intensity data in a case where the light intensity data exceeds a threshold value, the processing section processes the plurality of event data items excluding the marked light intensity data in accordance with an instruction to exclude the marked light intensity data, and a screening section is further provided to screen the particles associated with the plurality of event data items excluding the event data including the marked light intensity data in accordance with the instruction to exclude the marked light intensity data.

17. An information processing method comprising: a step of storing event data including light intensity data obtained by irradiating one of a plurality of particles with light; and a step of processing a plurality of event data items acquired from the plurality of particles, wherein the step of storing stores a mark assigned to the light intensity data in a case where the light intensity data exceeds a threshold value, and the step of processing processes the plurality of event data items excluding the marked light intensity data in accordance with an instruction to exclude the marked light intensity data.

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