Biological particle isolating device and biological particle isolating method

Abstract

The objective of the present disclosure is to satisfy the needs described in the specification. For example, one objective of the present disclosure is to provide technology that makes it possible to simultaneously obtain two specific types of isolated products from one sample.　The present disclosure provides a biological particle isolating device including: a determining unit that, for particles contained in a biological sample, performs isolation determination of the particles such that particles to be isolated and particles that are not to be isolated but are permitted to be isolated are isolated into a first accommodating space; and a display unit that displays information relating to the purity of the particles to be isolated contained in the first accommodation space. The present disclosure also provides a biological particle isolating method that includes: for particles contained in a biological sample, performing isolation determination of the particles such that particles to be isolated and particles that are not to be isolated but are permitted to be isolated are isolated into a first accommodating space; and displaying information relating to the purity of the particles to be isolated contained in the first accommodation space.

Description

Bioparticle sorting device and bioparticle sorting method

[0001] The present disclosure relates to a biological particle sorting device and a biological particle sorting method, and more particularly to a biological particle sorting device configured to perform sorting determination regarding particles to be sorted and particles that are not to be sorted but for which sorting is permitted, and a biological particle sorting method that includes performing the sorting determination.

[0002] For example, particle characteristics are measured by labeling a particle population, such as cells, microorganisms, and liposomes, with a fluorescent dye, irradiating each particle in the particle population with a laser beam, and measuring the intensity and / or pattern of fluorescence emitted from the excited fluorescent dye. An example of a biological sample analyzer that performs such measurements is a flow cytometer. Another example of a biological sample analyzer configured to separate cells is a cell sorter.

[0003] Flow cytometers and cell sorters can be configured to analyze multiple particles one by one by irradiating particles flowing in a single file through a flow channel with laser light (excitation light) of a specific wavelength and detecting the fluorescence and / or scattered light emitted from each particle. These devices convert the light detected by a photodetector into an electrical signal, which is then digitized and subjected to statistical analysis to determine the properties of each particle, such as type, size, and structure.

[0004] Several proposals have been made regarding the techniques for sorting performed in such biological sample analyzers. For example, Patent Document 1 listed below discloses a microparticle sorting device (claim 1) that includes a determination unit that performs particle sorting determination, the determination unit making the determination using rule data that defines the relationship between a particle population to which a particle to be sorted belongs and a particle population to which other particles within a predetermined range around the particle belong, and the particle populations to which the particle to be sorted and the other particles within the predetermined range may belong include the following particle populations (a) to (c), in which (a) a particle population of particles to be sorted, (b) a particle population of particles that are not to be sorted but can be ignored in the determination, and (c) a particle population of particles that are neither the particles to be sorted nor the negligible particles, and the negligible particles include red blood cells. The document states that the microparticle sorting device is equipped with a judgment unit that performs sorting judgment using the rule data, and therefore can perform sorting judgment that increases the recovery rate of particles to be sorted while reducing the impact on the purity of the particles to be sorted (paragraph 0018).

[0005] Japanese Patent Application Laid-Open No. 2022-17705

[0006] In a sorting process using a bioparticle sorting device such as a cell sorter, it is sometimes necessary to sort target cells with as high a purity as possible. Furthermore, when the number of target cells is small, it may be necessary to prioritize sorting efficiency so that as many target cells as possible are sorted, even if this results in a slight decrease in purity. Depending on the type or characteristics of the sample to be sorted using the device, it may be particularly desirable to address the latter need. Furthermore, it may be necessary to simultaneously meet both of these needs.

[0007] As described above, an example of a case where separation efficiency is prioritized is when a large number of particles that are acceptable for separation but are not the target for separation (also referred to herein as "particles that are asymmetric for separation and acceptable for separation") are present in the sample. Furthermore, it would be particularly useful if both of the above needs could be met simultaneously for the sample.

[0008] Therefore, an object of the present disclosure is to satisfy at least one of the above-mentioned needs, for example, to provide a particle sorting technique that is particularly suitable for biological samples that are not subject to sorting but contain a large number of particles that are permissible for sorting.

[0009] The present disclosure provides a biological particle sorting device having a determination unit that performs particle sorting determination for particles contained in a biological sample, such that particles to be sorted and particles that are not to be sorted but are acceptable for sorting are sorted into a first storage space, and a display unit that displays information regarding the purity of the particles to be sorted contained in the first storage space.The present disclosure also provides a biological particle sorting method, including: performing particle sorting determination for particles contained in a biological sample, such that particles to be sorted and particles that are not to be sorted but are acceptable for sorting are sorted into the first storage space, and displaying information regarding the purity of the particles to be sorted contained in the first storage space.

[0010] FIG. 1 is a schematic diagram showing an example of the configuration of a bioparticle sorting device according to the present disclosure. FIG. 1 is a schematic diagram showing an example of the configuration of a bioparticle sorting device according to the present disclosure. FIG. 2 is an example of a flow diagram of a bioparticle sorting operation performed by a bioparticle sorting device according to the present disclosure. FIG. 3 is a schematic diagram for explaining a sorted product. FIG. 4 is a schematic diagram showing an example of a screen on which a particle sorting mode is set. FIG. 5 is a schematic diagram showing an example of a screen on which a particle sorting mode is set. FIG. 6 is a schematic diagram showing an example of a screen on which a particle sorting mode is set. FIG. 7 is a schematic diagram showing an example of a screen on which a particle sorting mode is set. FIG. 8 is a schematic diagram showing an example of a screen on which a particle sorting mode is set. FIG. 9 is a schematic diagram showing an example of a screen on which a particle sorting mode is set. FIG. 10 is a schematic diagram showing an example of a screen on which a particle sorting mode is set. FIG. 11 is a schematic diagram showing an example of a screen on which a particle sorting mode is set. FIG. 12 is a schematic diagram showing an example of a screen showing the status of a particle sorting operation. FIG. 13 is a schematic diagram showing an example of a screen prompting selection of a particle sorting mode.

[0011] Preferred embodiments for carrying out the present disclosure will be described below. Note that the embodiments described below are representative embodiments of the present disclosure, and the scope of the present disclosure is not limited to these embodiments. Note that the present disclosure will be described in the following order: 1. First embodiment (bioparticle sorting device) (1) Basic concept (2) Configuration example (3) Example of sorting operation (4) Example of particle number counting (5) Example of screen for setting sorting mode (6) Example of screen for prompting switching to sorting mode 2. Second embodiment (bioparticle sorting method)

[0012] 1. First embodiment (bioparticle sorting device)

[0013] (1) Basic Concept The present disclosure relates to a biological particle sorting device. In one embodiment of the present disclosure, the biological particle sorting device may include a determination unit that performs particle sorting determination for particles contained in a biological sample, such that particles that are target for sorting and particles that are not target for sorting but are acceptable for sorting are sorted into a first storage space, and a display unit that displays information regarding the purity of the particles that are target for sorting contained in the first storage space. For example, the determination unit may perform the sorting determination in a sorting mode in which particles contained in the biological sample are sorted into two or more storage spaces, including the first storage space and a second storage space. The sorting mode may be configured to perform at least one of the following sorting operations (A) and (B): (A) a sorting operation in which only particles that are target for sorting are sorted into the second storage space; and (B) a sorting operation in which only particles that are target for sorting and particles that are not target for sorting but are acceptable for sorting are sorted into the first storage space. Furthermore, for example, a bioparticle sorting device according to the present disclosure may be configured to perform a particle sorting operation in a sorting mode in which two or more sorted products are obtained from a single biological sample. In the above sorting mode, at least two sorted products having different purities of particles to be sorted may be obtained. A bioparticle sorting device according to the present disclosure enables a sorting operation according to the properties of the sample and the needs of the user. For example, the bioparticle sorting device is suitable for performing a particle sorting operation on a biological sample containing a large number of particles that are not to be sorted but are acceptable for sorting. Furthermore, the present disclosure enables a particle sorting operation in which, for example, two sorted products having different purities of particles to be sorted from a single biological sample are obtained. Furthermore, a bioparticle sorting device according to the present disclosure can present useful information about the sorted products obtained by the sorting operation to the user.

[0014] For example, a particle sorting operation using a biological particle sorting device according to the present disclosure may be used when the proportion of target particles in a sample is very low. In this case, obtaining a sorted product recovered by prioritizing the purity of target particles and a sorted product recovered by prioritizing sorting efficiency, even though the purity of the target particles may be somewhat lower, would be beneficial for the user's analysis or development. For example, the sorted product recovered by prioritizing purity can be used to analyze only the target cells. Furthermore, the sorted product recovered by prioritizing sorting efficiency can be subjected to further sorting operations or used for analysis. Furthermore, when a particle sorting operation is performed on a biological sample containing non-target particles, such as red blood cells, if the non-target particles, such as red blood cells, are close to target particles, the target particles may be aborted, resulting in a reduced yield. By applying the particle sorting technique according to the present disclosure, such aborts can be avoided and the yield of target particles can be improved.

[0015] The information regarding the purity of the particles to be sorted contained in the first storage space may include at least one of the following: a ratio of the number of particles belonging to particle group (a) to the total number of particles belonging to particle groups (a) to (c) in the first storage space; a ratio of the number of particles belonging to particle group (a) to the total number of particles belonging to particle groups (a) and (b) in the first storage space; and a ratio of the number of particles belonging to particle group (a) to the total number of particles belonging to particle groups (a) and (c) in the first storage space. The biological particle sorting device (particularly the display unit) may be configured to display one or more of these ratios. Furthermore, the display unit may display information regarding the purity of the particles to be sorted contained in the second storage space. The information regarding the purity of the particles to be sorted contained in the second storage space may include at least one of the following: a ratio of the number of particles belonging to particle group (a) to the total number of particles belonging to particle group (a) below (a) to (c) below in the second storage space; a ratio of the number of particles belonging to particle group (a) to the total number of particles belonging to particle group (a) below (b) below in the second storage space; and a ratio of the number of particles belonging to particle group (a) to the total number of particles belonging to particle group (a) below (c) below in the second storage space. The biological particle sorting device (particularly the display unit) may be configured to display one or more of these ratios. This information is useful to a user performing a particle sorting operation. In some embodiments, the biological particle sorting device may be configured to display the ratio of the number of particles belonging to particle group (a) to the total number of particles belonging to particle group (a) below (b) below in the storage space. For example, the biological particle sorting device (particularly the display unit) may be configured to display the ratio of the number of particles belonging to particle group (a) to the total number of particles belonging to particle groups (a) and (b) in the first storage space. The biological particle sorting device can acquire the ratio as it performs the sorting operation described above. The ratio can be presented to a user as information regarding purity.

[0016] As described above, the determination unit may perform the sorting determination on particles contained in the biological sample in a sorting mode in which particles are sorted into two or more storage spaces including the first storage space and the second storage space, and may be configured to be able to perform at least one of the following sorting operations (A) and (B) in the sorting mode: (A) a sorting operation in which only particles to be sorted are sorted into the second storage space; and (B) a sorting operation in which only particles to be sorted and particles that are not to be sorted but are allowed to be sorted are sorted into the first storage space. In some embodiments, the biological particle sorting device may be configured to perform the following sorting operations (A') and (B') in the particle sorting operation in the sorting mode: (A') a sorting operation in which only particles to be sorted are sorted into one storage space; and (B') a sorting operation in which only particles to be sorted and particles that are not to be sorted but are allowed to be sorted are sorted into another storage space. The sorted product obtained in the second storage space (or one of the storage spaces) by the sorting operation of (A) (or (A')) and the sorted product obtained in the first storage space (or one of the other storage spaces) by the sorting operation of (B) (or (B')) may constitute two types of sorted products having different purities. The sorted product obtained in the second storage space (or one of the storage spaces) by the sorting operation of (A) (or (A')) has a high purity of particles to be sorted. Furthermore, the sorted product obtained in the first storage space (or one of the other storage spaces) by the sorting operation of (B) (or (B')) has a lower purity than the sorted product by (A) (or (A')), but the particles to be sorted can be efficiently recovered by the sorting operation of (B) (or (B')). Being configured to be able to perform the sorting operations (A) and (B) (or the sorting operations (A') and (B')) is particularly useful, for example, when the proportion of particles to be sorted out of all particles contained in the sample is low and the proportion of particles that are not to be sorted out but are acceptable to be sorted out is high.An example of such a case is when a blood-derived sample contains a low percentage of specific white blood cells, which are particles to be separated, and a large number of red blood cells, which are not the target particles but are acceptable for separation. In this case, the separation operation (A) (or (A')) can obtain a separated product containing the specific white blood cells at a high purity, and the separation operation (B) (or (B')) can obtain a separated product containing the specific white blood cells at a lower purity than the separated product obtained by the separation operation (A) (or (A')), but in which the specific white blood cells are recovered with a high separation efficiency. The separated product obtained by the separation operation (B) (or (B')) may contain red blood cells, but can be used in analysis or development where the presence of red blood cells is acceptable.

[0017] Another example of a case where the proportion of sorting target particles among all particles contained in a sample is low and the proportion of particles that are not sorted and are acceptable for sorting among all particles is high is when the sample contains, in addition to the sorting target cells, non-biological particles (e.g., beads) used for analyzing the sorting target cells. The non-biological particles may be, for example, non-biological particles used to capture the sorting target cells or non-biological particles used for analyzing specific components (e.g., analysis of components generated from the sorting target cells). In such a case, the sorting operation (A) (or (A')) can obtain a sorted product with a high purity of the sorting target cells, and the sorting operation (B) (or (B')) can obtain a sorted product in which the purity of the sorting target cells is lower than that of the sorting product obtained by the sorting operation (A) (or (A')), but in which the sorting target cells are recovered with high sorting efficiency. The fractionated product obtained by the fractionation operation (B) (or (B')) may contain non-biological particles, but can be used in analyses or developments where the presence of non-biological particles is acceptable, for example, when the non-biological particles can be easily removed. As described later, the particles may be biological particles such as cells and non-cellular biological particles, or non-biological particles such as beads. Furthermore, the beads may be beads having biological components (e.g., cells or cell-derived components, secretions, etc.) captured on their surface or lumen. The beads may be beads used for secretion analysis, for example. The particles may be collected in a state where they are contained within emulsion particles. In this case, the fractionated product may be an emulsion, and the dispersoid constituting the emulsion may be emulsion particles containing the particles to be fractionated. The dispersion medium constituting the emulsion may be appropriately selected by those skilled in the art depending on, for example, the type of emulsion particles. That is, the biological particle sorting device according to the present disclosure may be configured as a sorting device for biological component capturing particles (particularly biological component capturing beads), or may be configured as a sorting device for emulsion particles.In this way, the biological particle sorting device according to the present disclosure may be configured as a sorting device for particles other than biological particles (for example, beads or emulsion particles).

[0018] In order to perform the sorting operations (A) and (B) above (or the sorting operations (A') and (B')), for example, a determination unit that performs particle sorting determination may be used. That is, the biological particle sorting device may be equipped with a determination unit that performs particle sorting determination, and the determination unit may be configured to determine to which of the following particle populations (a) to (c) a particle to be determined and particles that may be present within a predetermined range around the particle to be determined belong, in a particle sorting operation in a sorting mode according to the present disclosure: (a) a particle population of particles to be determined; (b) a particle population of particles that are not to be determined and for which sorting is permitted; and (c) a particle population of particles that are not to be determined and for which sorting is not permitted. The sorting operations (A) and (B) above (or the sorting operations (A') and (B') above) can be performed by determining to which of these (a) to (c) a particle to be determined and particles that may be present within a predetermined range around the particle to be determined belong. For example, the biological particle sorting device sorts the target particles into one storage space when the target particles belong to the particle population (a) and no particles belonging to the particle populations (b) or (c) are present within the predetermined range, and sorts the target particles into another storage space when the target particles belong to the particle population (a) and particles belonging to the particle population (b) are present within the predetermined range but no particles belonging to the particle population (c). By configuring the biological particle sorting device to perform these two patterns of sorting, for example, a first sorted product containing the target particles with a high purity can be obtained in one storage space, and a second sorted product containing the target particles and the particles acceptable for sorting, but with a slightly lower purity than the first sorted product, can be obtained in the other storage space. The second sorted product contains the particles acceptable for sorting, and by obtaining the second sorted product, the target particles can be efficiently recovered.

[0019] (2) Configuration Example A configuration example of a biological particle sorting device configured to be able to perform a sorting operation in a sorting mode according to the present disclosure is described below. The biological particle sorting device according to the present disclosure may be configured as the biological sample analyzing device described below. That is, the "biological sample analyzing device 6100" described below may be read as the "biological particle sorting device 6100." Furthermore, the determination unit that performs particle sorting determination may be configured as the information processing unit 6103 described below. That is, the "information processing unit 6103" described below may be read as the "determination unit 6103."

[0020] An example configuration of a biological sample analyzer according to the present disclosure is shown in FIG. 1. The biological sample analyzer 6100 shown in FIG. 1 includes a light irradiation unit 6101 that irradiates light onto a biological sample S flowing through a flow path C, a detection unit 6102 that detects light generated by irradiating the biological sample S with light, and an information processing unit 6103 that processes information related to the light detected by the detection unit. Examples of the biological sample analyzer 6100 include a flow cytometer and an imaging cytometer. The biological sample analyzer 6100 may also include a fractionation unit 6104 that separates specific biological particles P from within the biological sample. An example of a biological sample analyzer 6100 that includes the fractionation unit is a cell sorter.

[0021] (Biological Sample) The biological sample S may be a liquid sample containing biological particles. The biological particles may be, for example, cells or non-cellular biological particles. The cells may be living cells, and more specific examples include blood cells such as red blood cells and white blood cells, and reproductive cells such as sperm and fertilized eggs. The cells may be directly collected from a specimen such as whole blood, or may be cultured cells obtained after culturing. Examples of the non-cellular biological particles include extracellular vesicles, particularly exosomes and microvesicles. The biological particles may be labeled with one or more labeling substances (e.g., dyes (particularly fluorescent dyes) and fluorescent dye-labeled antibodies). Note that the biological sample analyzer of the present disclosure may also analyze particles other than biological particles, such as beads for calibration purposes.

[0022] (Flow Channel) The flow channel C is configured to allow the biological sample S to flow. In particular, the flow channel C can be configured to form a flow in which biological particles contained in the biological sample are aligned in a substantially straight line. The flow channel structure including the flow channel C may be designed to form a laminar flow. In particular, the flow channel structure is designed to form a laminar flow in which the flow of the biological sample (sample flow) is surrounded by the flow of sheath liquid. The design of the flow channel structure may be appropriately selected by those skilled in the art, and a known design may be adopted. The flow channel C may be formed in a flow channel structure such as a microchip (a chip having flow channels on the order of micrometers) or a flow cell. The width of the flow channel C may be 1 mm or less, particularly 10 μm or more and 1 mm or less. The flow channel C and the flow channel structure including it may be formed from a material such as plastic or glass.

[0023] The biological sample analyzer of the present disclosure is configured so that light from light irradiation unit 6101 is irradiated onto the biological sample flowing within flow path C, and particularly onto biological particles within the biological sample. The biological sample analyzer of the present disclosure may be configured so that the interrogation point of light on the biological sample is within the flow path structure in which flow path C is formed, or so that the interrogation point of light is outside the flow path structure. An example of the former is a configuration in which the light is irradiated onto flow path C within a microchip or flow cell. In the latter, the light may be irradiated onto biological particles after they have left the flow path structure (particularly its nozzle portion), and an example of this is a jet-in-air flow cytometer.

[0024] (Light Irradiation Unit) The light irradiation unit 6101 includes a light source unit that emits light and a light-guiding optical system that guides the light to an irradiation point. The light source unit includes one or more light sources. The type of light source is, for example, a laser light source or an LED. The wavelength of the light emitted from each light source may be any of ultraviolet light, visible light, and infrared light. The light-guiding optical system includes optical components such as a beam splitter group, a mirror group, or an optical fiber. The light-guiding optical system may also include a lens group for focusing light, such as an objective lens. There may be one or more irradiation points where the light intersects with the biological sample. The light irradiation unit 6101 may be configured to focus light irradiated from one or more different light sources onto one irradiation point.

[0025] (Detection Unit) The detection unit 6102 includes at least one photodetector that detects light generated by irradiating the bioparticles with light. The detected light is, for example, fluorescence or scattered light (e.g., one or more of forward scattered light, back scattered light, and side scattered light). Each photodetector includes one or more light-receiving elements, for example, a photodetector array. Each photodetector may include one or more PMTs (photomultiplier tubes) and / or photodiodes such as APDs and MPPCs as light-receiving elements. The photodetector includes, for example, a PMT array in which multiple PMTs are arranged in a one-dimensional direction. The detection unit 6102 may also include an imaging element such as a CCD or CMOS. The detection unit 6102 can acquire images of the bioparticles (e.g., bright-field images, dark-field images, and fluorescence images) using the imaging element.

[0026] The detection unit 6102 includes a detection optical system that allows light of a predetermined detection wavelength to reach a corresponding photodetector. The detection optical system includes a spectroscopic unit such as a prism or a diffraction grating, or a wavelength separation unit such as a dichroic mirror or an optical filter. The detection optical system is configured to, for example, disperse light generated by irradiating bioparticles with light, and detect the dispersed light using a plurality of photodetectors, the number of which is greater than the number of fluorescent dyes with which the bioparticles are labeled. A flow cytometer that includes such a detection optical system is called a spectral flow cytometer. The detection optical system is also configured to, for example, separate light corresponding to the fluorescent wavelength range of a specific fluorescent dye from the light generated by irradiating bioparticles with light, and detect the separated light using a corresponding photodetector.

[0027] The detection unit 6102 may also include a signal processing unit that converts the electrical signal obtained by the photodetector into a digital signal. The signal processing unit may include an A / D converter as a device that performs the conversion. The digital signal obtained by the conversion by the signal processing unit may be transmitted to the information processing unit 6103. The digital signal may be handled by the information processing unit 6103 as data related to light (hereinafter also referred to as "light data"). The light data may be light data including, for example, fluorescent light data. More specifically, the light data may be light intensity data, and the light intensity may be light intensity data of light including fluorescent light (which may include feature quantities such as area, height, and width).

[0028] (Information Processing Unit) The information processing unit 6103 includes, for example, a processing unit that processes various data (e.g., optical data) and a storage unit that stores various data. When the processing unit acquires optical data corresponding to a fluorescent dye from the detection unit 6102, the processing unit may perform fluorescence spillover correction (compensation processing) on ​​the light intensity data. Furthermore, in the case of a spectral flow cytometer, the processing unit performs fluorescence separation processing on the optical data to acquire light intensity data corresponding to the fluorescent dye. The fluorescence separation processing may be performed, for example, according to the unmixing method described in Japanese Patent Application Laid-Open No. 2011-232259. When the detection unit 6102 includes an image sensor, the processing unit may acquire morphological information of bioparticles based on images acquired by the image sensor. The storage unit may be configured to store the acquired optical data. The storage unit may further be configured to store spectral reference data used in the unmixing processing.

[0029] If the biological sample analyzer 6100 includes a fractionating unit 6104 (described below), the information processing unit 6103 can determine whether to fractionate bioparticles based on the optical data and / or morphological information. The information processing unit 6103 can then control the fractionating unit 6104 based on the result of this determination, allowing the fractionating unit 6104 to fractionate the bioparticles.

[0030] The information processing unit 6103 may be configured to output various data (e.g., optical data, images, etc.). For example, the information processing unit 6103 may output various data (e.g., two-dimensional plots, spectral plots, etc.) generated based on the optical data. The information processing unit 6103 may also be configured to accept input of various data, such as accepting gating processing on a plot by a user. The information processing unit 6103 may include an output unit (e.g., a display, etc.) or an input unit (e.g., a keyboard, etc.) for executing the output or input.

[0031] The information processing unit 6103 may be configured as a general-purpose computer, for example, as an information processing device including a CPU, RAM, and ROM. The information processing unit 6103 may be included in a housing that includes the light irradiation unit 6101 and the detection unit 6102, or may be located outside the housing. Furthermore, various processes or functions performed by the information processing unit 6103 may be realized by a server computer or a cloud connected via a network.

[0032] (Sorting unit) The sorting unit 6104 sorts the bioparticles according to the determination result by the information processing unit 6103. The sorting method may be a method of generating droplets containing bioparticles by vibration, applying an electric charge to the droplets to be sorted, and controlling the direction of travel of the droplets using electrodes. The sorting method may also be a method of controlling the direction of travel of the bioparticles within the flow channel structure to perform sorting. The flow channel structure is provided with, for example, a control mechanism using pressure (spray or suction) or electric charge. An example of such a flow channel structure is a chip (for example, the chip described in JP 2020-76736 A) having a flow channel structure in which a flow channel C branches downstream into a recovery flow channel and a waste flow channel, and in which specific bioparticles are recovered into the recovery flow channel.

[0033] (3) Example of Sorting Operation An example of a sorting operation according to the present disclosure will be described below with reference to Figures 2, 3A, and 3B. Figure 2 is a diagram showing an example of the configuration of a bioparticle sorting device according to the present disclosure. Figure 3A is a flow diagram of a sorting process performed by the bioparticle sorting device. Figure 3B is a schematic diagram for explaining a sorted product obtained by the sorting process.

[0034] (Device Configuration Example) The bioparticle sorting device 100 shown in FIG. 2 includes a light irradiation unit 101, a detection unit 102, and an information processing unit (determination unit) 103. Examples of the bioparticle sorting device 100 include a cell sorter, a flow cytometer, and an imaging cytometer. The bioparticle sorting device 100 further includes a sorting unit 104 that sorts the bioparticles. The light irradiation unit 101, the detection unit 102, the information processing unit 103, and the sorting unit 104 correspond to the light irradiation unit 6101, the detection unit 6102, the information processing unit 6103, and the sorting unit 6104 described in (2) above. Therefore, the explanations regarding the light irradiation unit 6101, the detection unit 6102, the information processing unit 6103, and the sorting unit 6104 also apply to the light irradiation unit 101, the detection unit 102, the information processing unit 103, and the sorting unit 104. As described below, the sorting unit 104 may be configured to sort bioparticles in an open space or a closed space. That is, the bioparticle sorting device may be configured as a flow cytometer that sorts bioparticles in an open space or a closed space.

[0035] In addition, the bioparticle sorting device 100 may further include a liquid delivery unit 112 that supplies various liquids (e.g., sample liquid and sheath liquid) to a chip 111 having a flow path through which the sample flows, a memory unit 113, a display unit 114, and a user interface (UI) 115.

[0036] The chip 111 may be, for example, a microchannel chip. Alternatively, a flow cell may be used instead of the chip. The chip 111 may be replaceably attached to the bioparticle sorting device 100. The structure of the flow channel P of the chip 111 may be designed as appropriate by a person skilled in the art, and in particular may be configured to form a laminar flow composed of a sample liquid, which is a specimen containing bioparticles, and a sheath liquid.

[0037] The liquid delivery unit 112 delivers a sample liquid, which is a specimen containing bioparticles, to the chip and / or delivers a sheath liquid to the chip. The liquid delivery unit may include, for example, a pump. In the chip shown in FIG. 2 , the liquid delivery unit introduces the sample liquid into the sample liquid flow path P11 and the sheath liquid into two sheath liquid flow paths P12a and P12b. The sample liquid flow path P11 and the sheath liquid flow paths P12a and P12b merge to form a main flow path P13. The sample liquid laminar flow delivered through the sample liquid flow path P11 and the sheath liquid laminar flow delivered through the sheath liquid flow paths P12a and P12b merge in the main flow path P13, forming a sheath flow in which the sample liquid laminar flow is sandwiched between the sheath liquid laminar flows.

[0038] The storage unit 113 can store various data, such as various programs used to control the bioparticle sorting device, data related to the results of sorting determination, and data related to the number of particles. The display unit 114 can display various screens used by the user to use the bioparticle sorting device. The UI 115 can include, for example, various input / output devices. The user can operate the bioparticle sorting device via the UI.

[0039] (Processing Flow) An example of the processing flow of the sorting operation performed by the bioparticle sorting device 100 will be described with reference to Figure 3. This processing flow is an example of a processing flow from sorting determination based on the detection result of light generated by irradiating light onto one particle to be determined to the sorting operation. Note that the light irradiation may be performed by the light irradiation unit. Furthermore, the light detection may be performed by the detection unit.

[0040] When the biological particle sorting device 100 (A) executes a sorting operation in which only particles to be sorted are sorted into one storage space, the particles to be sorted are recovered into the second container 122 by the sorting operation. When the biological particle sorting device 100 (B) executes a sorting operation in which only particles to be sorted and particles that are not to be sorted and are permissible for sorting are sorted into other storage spaces, the particles to be sorted are recovered into the first container 121 by the sorting operation. The sorting operation (B) can be executed, for example, when only particles that are not to be sorted and are permissible for sorting are present near the particles to be sorted. When these sorting operations are not executed, the particles to be determined may be recovered into the third container 123. Alternatively, in this case, the particles to be determined may be recovered into another flow path.

[0041] In step S11, the biological particle sorting device 100 determines whether the particle to be determined is a particle to be sorted. If the particle to be determined is a particle to be sorted, the biological particle sorting device 100 proceeds to step S12. If the particle to be determined is not a particle to be sorted, the biological particle sorting device 100 proceeds to step S13. Step S11 may be executed by the information processing unit 103 in particular.

[0042] In step S12, the biological particle sorting device 100 determines to which of the following particle populations (a) to (c) particles each of which may be present within a predetermined range around the target particle (the target particle for sorting) belongs: (a) a particle population of particles to be sorted; (b) a particle population of particles that are not to be sorted but are permissible for sorting; and (c) a particle population of particles that are not to be sorted and are not permissible for sorting. Note that the particles contained in the biological sample may belong to any of these (a) to (c). For example, the particles in the biological sample may be composed of particles belonging to the particle population (a), particles belonging to the particle population (b), and other particles (corresponding to particles belonging to the particle population (c)).

[0043] If the bioparticle sorting device 100 determines that no particles belonging to the particle group (b) or (c) are present within the predetermined range (determined as determination result A in the figure), the process proceeds to step S13. The bioparticle sorting device 100 may also proceed to step S13 if particles belonging to the particle group (a) are present within the predetermined range (i.e., if one or more particles to be sorted are present within the predetermined range around the particle to be sorted). If the bioparticle sorting device 100 determines that particles belonging to the particle group (b) are present within the predetermined range but no particles belonging to the particle group (c) are present (determined as determination result B in the figure), the process proceeds to step S14. If the bioparticle sorting device 100 determines that particles belonging to the particle group (c) are present within the predetermined range (determined as determination result C in the figure), the process proceeds to step S15. Note that even if particles belonging to the particle group (b) are present within the predetermined range, if it is determined that particles belonging to the particle group (c) are present within the predetermined range, the biological particle sorting device 100 proceeds to step S15. As will be described later, different operations are performed in steps S13, S14, and S15. That is, the biological particle sorting device 100 selects an operation to be performed depending on the type of particles present within the predetermined range. Step S12 may be performed by the information processing unit 103 in particular.

[0044] The bioparticle sorting device can determine, for example, whether particles exist within a predetermined range around the target particle based on the time when light generated by irradiating the target particle with light is detected and the time when light generated by irradiating other particles flowing in front of and / or behind the target particle is detected. The velocity of these particles within the flow path C can be calculated, for example, based on the difference between the detection time of light generated by irradiating the particle with light at one position and the detection time of light generated by irradiating the particle with light at another position. For this calculation, the distance between the two irradiation positions may be determined in advance, and the velocity of the microparticle can be determined based on the difference between the two detection times and the distance. The bioparticle sorting device can determine whether other particles exist within the predetermined range based on the difference between the detection time of light generated by irradiating the target particle with light and the detection time of light generated by irradiating the other particles with light. More specifically, the bioparticle sorting device can determine whether other particles exist within the predetermined range based on whether the difference is equal to or less than a predetermined value. In this manner, the predetermined range may be a range determined based on the predetermined value relating to the time difference. The predetermined value can be determined in advance. For example, the predetermined value can be determined by determining how far other particles must be from the target particle to prevent the other particles from being contained in the droplet when the droplet is formed. Note that, in this specification, the predetermined value is also referred to as a "guard time." Furthermore, since the predetermined range can be determined based on the predetermined value as described above, the term "guard time" is sometimes used to refer to the predetermined range. Furthermore, although the above describes a method for determining whether a particle exists within a predetermined range based on the particle's velocity, other methods may also be used in the present disclosure to determine whether a particle exists within a predetermined range.

[0045] As described above, the biological particle sorting device can determine whether to collect a particle to be determined based on whether other particles are present within a predetermined range around the particle to be determined and / or the type of the other particles. If other particles are present within the predetermined range, a determination is made not to collect the particle to be determined, thereby increasing the purity of the target particle among the collected particles, as described above. On the other hand, if other particles are present within the predetermined range and their inclusion in the sorted product is acceptable (e.g., if the other particles do not adversely affect the analysis or use of the sorted product), collecting the particle to be determined also collects the other particles in the same container. Therefore, although the purity of the particle to be determined in the sorted product is somewhat reduced, the recovery rate of the particle to be determined can be increased.

[0046] In step S13, the biological particle sorting device 100 executes a sorting operation to sort the target particles (which are sorting target particles) into the second container 122. Since it has been determined that no particles belonging to the particle populations (b) or (c) are present within the predetermined range around the target particles, the sorting operation is executed to collect the target particles into the second container 122. Therefore, the sorted product obtained in the second container by repeating step S13 is a sorted product containing the target particles with extremely high purity. The purity may be, for example, 90% or more, particularly 93% or more, more particularly 95% or more, even more particularly 97% or more, 98% or more, or 99% or more. The purity is the ratio of the number of target particles to the total number of particles in the sorted product. Note that the sorted product may contain particles other than the target particles, and the purity may be 100% or less. In step S13, the sorting unit 104 of the bioparticle sorting device 100, in particular, executes a sorting operation. As described in (2) above, the sorting operation may be an operation in which the sorting unit 104 generates droplets containing particles to be sorted by vibration, applies an electric charge to the droplets, and controls the direction of the droplets with electrodes, thereby causing the droplets to travel toward the second container 122. In this way, the sorting operation may be a sorting operation in which bioparticles are sorted in an open space. Alternatively, the sorting operation may be another sorting operation described above, or may be a sorting operation in a closed space, such as a sorting operation by suction. In this way, in step S13, a sorting operation in which only the particles to be sorted are sorted into one storage space may be executed.

[0047] In step S14, the biological particle sorting device 100 executes a sorting operation to sort the target particles (i.e., particles to be sorted) into the first container 121. Since particles belonging to the particle group (b) are present within the predetermined range around the target particles, but particles belonging to the particle group (c) are not present, by executing the sorting operation, particles belonging to the particle group (b) are collected into the first container 121 in addition to the target particles. Therefore, the sorted product obtained by repeating step S14 is a sorted product that includes not only the target particles but also particles that are not to be sorted but are permissible for sorting. The purity of the target particles in the sorted product may be lower than the purity of the target particles in the sorted product obtained by repeating step S13, for example, 70% or less, 65% or less, or 60% or less. Furthermore, the purity of the sorted product obtained by repeating step S14 may be, for example, 30% or more, 35% or more, or 40% or more. When the content of the allowable particles among particles contained in a biological sample is high and the proportion of the target particles is very small, attempting to separate only the target particles may result in a decrease in the efficiency of separation of the target particles due to the large number of allowable particles. According to the present disclosure, as described above, by separating the target particles even when the allowable particles are within the predetermined range, the efficiency of separation of the target particles can be improved. In step S14, the sorting unit 104 of the biological particle sorting device 100, in particular, performs the sorting operation. As described in (2) above, the sorting operation may be an operation in which the sorting unit 104 generates droplets containing the target particles by vibration, applies an electric charge to the droplets, and controls the direction of travel of the droplets with electrodes to cause the droplets to travel toward the first container 121. In this manner, the sorting operation may be an operation in which biological particles are separated in an open space. Alternatively, the sorting operation may be another sorting operation described above, or may be a sorting operation in a closed space, such as a sorting operation by suction, etc. In this way, in step S14, a sorting operation may be performed in which only the particles to be sorted and particles that are not to be sorted but are allowed to be sorted are sorted into another storage space.

[0048] In step S15, the bioparticle sorting device 100 collects the target particles into the third container 123. Since particles belonging to the particle group (c) are present within the predetermined range around the target particle, the target particles are collected into the third container 123, and the particles belonging to the particle group (c) are also collected into the third container 123. In step S15, the sorting unit 104 of the bioparticle sorting device 100, in particular, executes a collecting operation. This collecting operation may be, for example, an operation in which the generated droplets are allowed to travel toward the third container 123 without controlling the direction of travel of the droplets using electrodes, as described in (2) above. The product in the third container may be treated as waste liquid, or may be used as a sample for other target particles.

[0049] In step S16, the bioparticle sorting device 100 collects the particles to be determined into the third container 123. In step S16, the sorting unit 104 of the bioparticle sorting device 100 executes the collecting operation. This collecting operation may be, for example, an operation in which the generated droplets are allowed to travel toward the third container 123 without controlling the direction of travel of the droplets using electrodes, as described in (2) above. The product in the third container may be treated as waste liquid, or may be used as a sample for other particles to be sorted.

[0050] By performing the above-described process flow, a fractionated product containing highly purified particles to be separated is formed in the second container, and a fractionated product containing relatively high purity particles to be separated is formed in the first container, although the purity is lower than that of the fractionated product in the second container. Figure 3B is a schematic diagram showing only the portion S enclosed by the dashed line in Figure 2. As shown in Figure 3B, a fractionated product containing highly purified particles to be separated 131 is obtained in the second container 122, and a fractionated product containing particles 132 that are not to be separated but are acceptable for separation, in addition to the particles to be separated 131, is obtained in the first container 121. The purity of the particles to be separated in the first container 121 is lower than the purity of the particles to be separated in the second container 122. Thus, the present disclosure makes it possible to obtain two or more fractionated products containing particles to be separated that differ in purity from one another from a single biological sample. Furthermore, the fractionated product in the first container includes not only particles to be separated but also particles that are not to be separated, but since the particles that are not to be separated are particles that are acceptable for separation, the fractionated product can be used in analyses in which the presence of such particles is acceptable. Furthermore, by obtaining the fractionated product in the first container in addition to the fractionated product in the second container, the yield of particles to be separated can be increased.

[0051] (4) Example of Particle Counting The biological particle sorting device according to the present disclosure may be configured to count the number of particles belonging to the particle group (b) described in (3) above, i.e., the number of particles that are not subject to sorting but are permissible for sorting. The number of particles may be the number of particles in any of the storage spaces (particularly any of the containers), and may be the number of particles in the first storage space. The counting may be performed, for example, by counting the number of times the information processing unit determines that a particle (e.g., a particle to be determined or particles surrounding a particle to be determined) belongs to the particle group (b). Note that, in this specification, the number of particles belonging to the particle group (a), the number of particles belonging to the particle group (b), and the number of particles belonging to the particle group (c) are also referred to as the "number of (a)," the "number of (b)," and the "number of (c)," respectively. Furthermore, these numbers may be the number of particles in any of the storage spaces (particularly any of the containers).

[0052] The number of (b) is useful for identifying the constituent ratio of particles in a sample. The number of (b) is also useful for identifying the recovery efficiency (also called "efficiency") of particles to be sorted. These are particularly useful when the constituent ratio of (b) in a sample is high.

[0053] A bioparticle sorting device according to the present disclosure may be configured to display the number (b), or a value or index calculated using the number (b). Examples of values ​​calculated using the number (b) include, but are not limited to, values ​​(particularly ratios) calculated using the number (b) as the numerator or one element of the numerator, or values ​​(particularly ratios) calculated using the number (b) as the denominator or one element of the denominator. Examples of values ​​calculated using the number (b) include the above-mentioned recovery efficiency of particles to be sorted. The value may be a ratio expressed by the following formula, for example: Ratio 1 = (a) / ((a)+(b)+(c)). In this formula, (a), (b), and (c) represent the number (a), the number (b), and the number (c), respectively. The same applies to the other formulas below. Alternatively, the value may be a ratio expressed by the following formula: Ratio 2 = (b) / ((a)+(b)+(c)). Alternatively, the value may be a ratio expressed by the following formula: Ratio 3 = ((a) + (b)) / ((a) + (b) + (c)) Examples of the index calculated using the number (b) include a determination result based on the number (b) or a determination result based on a value calculated using the number (b). For example, the determination result may be a display meaning that the recovery efficiency is good (e.g., letters or symbols indicating that the recovery efficiency is good, such as "Good") or a display meaning that the recovery efficiency is not good (e.g., letters or symbols indicating that the recovery efficiency is not good, such as "Not Good"). For example, such a determination result may be output when the value is equal to or greater than a predetermined threshold value or when it is less than the threshold value.

[0054] Of course, the bioparticle sorting device according to the present disclosure may display a value or index calculated without using the number (b). Furthermore, the bioparticle sorting device according to the present disclosure may display the number (a) and / or the number (c). Examples of values ​​calculated without using the number (b) include, but are not limited to, values ​​(particularly, ratios) calculated using the number (a) as a numerator or as one element of a numerator. Examples of values ​​calculated using the number (a) include the above-mentioned recovery efficiency of the particles to be sorted. This value may be, for example, a ratio expressed by the following formula: Ratio 4 = (a) / ((a) + (c)). Examples of indices calculated without using the number (b) include a determination result based on the number (a) or a determination result based on a value calculated using the number (a). For example, the determination result may be an indication that the recovery efficiency is good or poor. For example, when the recovery efficiency is equal to or greater than a predetermined threshold value or is less than a threshold value, such a determination result may be output.

[0055] Furthermore, the bioparticle sorting device according to the present disclosure may calculate the following ratio 5 and / or ratio 6. Ratio 5 = (a) / ((a) + (b)) Ratio 6 = (b) / ((a) + (b)) The bioparticle sorting device according to the present disclosure (particularly the display unit) may display one or more of the ratios described above (particularly ratios 1 to 6). The ratio may be the ratio within any of the storage spaces described above, and particularly may be the ratio within the first storage space. In one embodiment, the bioparticle sorting device (particularly the display unit) may display ratio 5. Ratio 5 may, for example, be the ratio within any of the storage spaces described above, and particularly may be the ratio within the first storage space. The bioparticle sorting device according to the present disclosure may have a display unit that displays information regarding the purity of the particles to be sorted contained in the first storage space, and the information may include these ratios.

[0056] Furthermore, the biological particle sorting device according to the present disclosure may calculate the following sum 1 and / or sum 2: Sum 1 = (a) + (b) Sum 2 = (a) + (b) + (c) The biological particle sorting device according to the present disclosure (particularly the display unit) may display one or more of the above-described sums (particularly sums 1 and 2). The sum may be the total number of particles in any of the above-described storage spaces, and may particularly be the proportion within the first storage space. The biological particle sorting device according to the present disclosure may have a display unit that displays information regarding the purity of the particles to be sorted contained in the first storage space, and the display unit may display the total number as part of the information regarding the purity or as information separate from the information regarding the purity.

[0057] In the present disclosure, the number of (b) counted per unit time may also be calculated. Similarly, the number of (a) counted per unit time or the number of (c) counted per unit time may also be calculated. The bioparticle sorting device according to the present disclosure (particularly the display unit) may display values ​​related to such event frequencies.

[0058] For example, (a) / ((a)+(c)) may be used as purity, but by using the number of (b), it is possible to know the extent to which (a) is contained in (a)+(b)+(c). In addition, the present disclosure also makes it possible to know the number of (b) itself. In the present disclosure, as an index for a sorted product intended to separate only particles belonging to the particle population of (a) (the sorted product collected in the first container described above), the ratio of particles to be sorted (particularly the ratio of cells to be sorted) for (a) may be expressed, for example, by (a) / ((a)+(b)). Furthermore, when a sorted product containing only particles belonging to the particle population of (a) and a sorted product containing only particles belonging to the particle population of (b) are obtained, (a) / ((a)+(c)) may be used as the purity of (a). In this case, the true ratio of (a) may be determined by presenting the ratio of (b).

[0059] By displaying the number of (b) or a value or index calculated using the number of (b), it is possible to provide the user with information useful for analyzing biological samples when the sample contains particles of (b), particularly when the sample contains a large number of particles belonging to the particle population of (b).

[0060] Furthermore, a predetermined particle sorting operation may be performed based on the number of (b). For example, the biological particle sorting device may be configured to perform a particle sorting operation in a sorting mode to obtain the two or more sorted products when the number of particles belonging to the particle population of (b) is equal to or greater than a predetermined value, or when the ratio of the number of particles belonging to the particle population of (b) to the number of particles belonging to the particle population of (a) is equal to or greater than a predetermined value. In this way, the biological particle sorting device according to the present disclosure may automatically adopt a specific sorting mode depending on the number of (b).

[0061] (5) Example of a screen for setting the sorting mode The biological particle sorting device according to the present disclosure may be configured to be able to perform particle sorting operations in other sorting modes in addition to the sorting mode for obtaining the two or more sorted products.

[0062] For example, the biological particle sorting device may execute a sorting process in which, when the determination result A described in (3) above is obtained for a particle to be determined, the particle to be determined is sorted, and, when the determination results B and C described in (3) above are obtained for a particle to be determined, the particle to be determined is not sorted and the process is aborted. In other words, the biological particle sorting device may be configured to execute a sorting mode in which a sorted product with high purity of the particle to be determined is obtained.

[0063] The sorting mode may be set by, for example, receiving a user input on a screen displayed on a display unit of the bioparticle sorting device.

[0064] In one embodiment, the bioparticle sorting device is configured to display a screen on a display unit prompting a user to select a sorting mode, and the screen may be configured to allow a user to select a sorting mode for obtaining the two or more sorted products. Examples of such screens will be described with reference to Figures 4A to 4H. Screen 200 shown in the figures is an example of a screen when the bioparticle sorting device is configured to collect the sorted products into each of four containers.

[0065] The biological particle sorting device (e.g., a display unit) displays a screen (e.g., a window) 200 shown in Fig. 4A. This screen may be displayed at any stage before a sorting operation on a biological sample is performed, for example, but may also be displayed at other stages. In screen 200, Far Left, Left, Right, and Far Right are displayed as the names of the four containers, respectively. These displays also indicate the positions of these containers.

[0066] As shown in FIG. 4B , the screen 200 has a sorting method selection field 201 (Sort Type), a sorting gate selection field 202 (Sort Gate), a sorting allowable gate selection field 203 (Ignore Gate), and a sorting stop number selection field 204 (Stop Count) associated with each of the four containers. The sorting allowable gate may also be called an ignore gate. Each selection field may be displayed as a drop-down box, as shown in the figure, for example, but may also be displayed in other formats. Each selection field will be described below.

[0067] When the user selects the sorting method selection field 201 to specify the sorted product to be formed into the Far Left container, a list of selectable sorting methods is displayed, as shown in FIG. 4C . In the same figure, the selectable sorting types, Purity and Ignore, are displayed in the list of selectable sorting methods. When the user selects the desired sorting method from these sorting methods, the sorting method for obtaining the sorted product to be formed into the Far Left container is specified.

[0068] Furthermore, when the user selects the sorting gate selection field 202 to specify the particles to be sorted to be included in the sorted product formed in the Far Left container, a list of selectable gates is displayed, as shown in FIG. 4D , for example. In the same figure, four gates A to D are displayed in the list as selectable gates. Each gate may be set in advance by the user. The gate setting may be performed by the user as appropriate, for example, depending on the particles to be sorted and / or the biological sample. When the user selects a desired gate from these sorting gates, the particles to be sorted to the Far Left container are specified.

[0069] Furthermore, when the user selects the sorting acceptable gate selection field 203 to specify particles that are acceptable to be included in the sorted product formed in the Far Left container but are not the target of sorting, a list of selectable gates is displayed, as shown in FIG. 4E, for example. In the same figure, four gates A to D are displayed in the list as selectable gates. Each gate may be set appropriately by the user. By selecting a sorting acceptable gate from among these gates, the user specifies particles that are acceptable to be sorted into the Far Left container but are not the target of sorting.

[0070] Furthermore, in order for the user to specify the number of particles to be collected into the Far Left container, the stop fraction count selection field 204 may be configured to allow the user to select a desired number from a list or to input a desired number. The stop fraction count may be such that the operation of collecting fractions into the container is stopped once the number of fractions selected or input in the field has been collected.

[0071] For example, depending on the sorting method selected in the sorting method selection field, the sorting acceptable gate selection field may be displayed as selectable or unselectable. For example, with respect to the purity priority method (Purity), the sorting acceptable gate selection field does not need to be considered. Therefore, the bioparticle sorting device may make the sorting acceptable gate selection field unselectable, for example, by disabling a drop-down box, in response to the purity priority method being selected in the sorting method selection field. An example of such an unselectable state is shown by reference numeral 208 in FIG. 4F . On the other hand, with the ignore method (Ignore), the sorting acceptable gate selection field needs to be considered. Therefore, the bioparticle sorting device may make the sorting acceptable gate selection field selectable, for example, by enabling a drop-down box, in response to the ignore method being selected in the sorting method selection field. In this way, the bioparticle sorting device may change the sorting acceptable gate selection field depending on the selected sorting method. This prevents the user from making unnecessary gate selections.

[0072] FIG. 4F shows the screen after configuration for four containers.

[0073] For the Far Left container, the purity-priority method is selected as the sorting method, gate A is selected as the sorting gate, and 10,000 is selected as the sorting stop number. For the Left container, the ignore method is selected as the sorting method, gate A is selected as the sorting gate, gate B is selected as the sorting allowable gate, and 1,000 is selected as the sorting stop number. That is, based on the settings for the Far Left container and the settings for the Left container, a sorting operation according to the present disclosure is performed on particles belonging to gate A, and two types of sorted products with different purities of sorting target particles (particles belonging to gate A) are obtained. Also, in the same figure, the purity-priority method and the ignore method are selected for the same sorting target particles (particles belonging to gate A). In this way, when both the purity-priority method and the ignore method are set for the same sorting target particles (e.g., particles belonging to the same gate), a sorting operation using the purity-priority method may be preferentially assigned to certain sorting target particles, for example.

[0074] Furthermore, for the Right container, the purity-priority method is selected as the sorting method, gate C is selected as the sorting gate, and 10,000 is selected as the sorting stop number. For the Far Right container, the ignore method is selected as the sorting method, gate C is selected as the sorting gate, gate D is selected as the sorting allowable gate, and 1,000 is selected as the sorting stop number. That is, based on the settings for the Right container and the settings for the Far Right container, a sorting operation according to the present disclosure is performed on particles belonging to gate C, and two types of sorted products with different purities of particles to be sorted (particles belonging to gate C) are obtained. Also, in the same figure, the purity-priority method and the ignore method are selected for the same particles to be sorted (particles belonging to gate C). As described above, when both the purity-priority method and the ignore method are set for the same particles to be sorted, for example, a sorting operation using the purity-priority method may be preferentially assigned to a certain particle to be sorted.

[0075] In this way, the bioparticle sorting device according to the present disclosure is equipped with a display unit that displays a screen that prompts the user to select a sorting mode, and the screen may be configured to allow the user to select a sorting mode for obtaining the two or more sorted products.

[0076] Furthermore, various particle sorting operations can be set using such a screen. For example, the biological particle sorting device can perform a particle sorting operation in a sorting mode in which a sorting operation is performed only in the purity-priority mode for each of two or more types of sorting target particles, thereby obtaining a sorted product with a high purity for each of the two or more types of sorting target particles. For example, the device may be configured to perform a sorting mode in which a sorted product with a high purity for sorting target particle A, a sorted product with a high purity for sorting target particle B, a sorted product with a high purity for sorting target particle C, and a sorted product with a high purity for sorting target particle D are obtained. An example of a screen set for performing a particle sorting operation in such a sorting mode is shown in FIG. 4G. As shown in the figure, the purity-priority mode may be selected as the sorting mode for each of sorting target particles A to D.

[0077] The biological particle sorting device may be configured to perform a particle sorting operation in a particle sorting mode that obtains two sorted products with different purities for each of one or more types of particles to be sorted according to the present disclosure. An example of a screen setting for performing the particle sorting operation is as described above in Fig. 4F, and the screen in Fig. 4F is configured to perform a particle sorting operation in a sorting mode that obtains two sorted products with different purities for each of two types of particles to be sorted (particles belonging to gate A and particles belonging to gate C).

[0078] The biological particle sorting device may also perform a particle sorting operation in a sorting mode to obtain a high-purity sorted product for each of one or more types of sorting target particles, and may also perform a particle sorting operation to obtain two sorted products with different purities for each of one or more other types of sorting target particles according to the present disclosure. An example of a screen set for performing such a particle sorting operation in a sorting mode is shown in Figure 4H. The screen in Figure 4H is set to perform a particle sorting operation in a sorting mode to obtain two sorted products with different purities for each of one type of sorting target particles (particles belonging to gate A), and the purity-priority method is selected as the sorting method for each of the two types of sorting target particles (particles belonging to gate C and particles belonging to gate D).

[0079] (6) Example of a Screen Prompting to Switch Sorting Modes A biological particle sorting device according to the present disclosure may be configured to allow the sorting mode to be switched during a particle sorting operation on a sample. For example, the proportion of particles to be sorted and / or the proportion of particles that are not to be sorted but are acceptable for sorting may only be determined after the particle sorting operation has been performed. By being able to change the sorting mode once these proportions are known, an appropriate sorting mode can be selected according to these proportions. In this way, by displaying a screen prompting the user to select a sorting mode during a particle sorting operation, particle sorting operations tailored to the user's needs can be performed.

[0080] Examples of the display aspects of the screen will be described with reference to Figures 5A to 5C. Figure 5A is an example of a screen showing the status of a sorting operation. The screen shown in the figure shows a situation in which sorting of particles belonging to gate A and sorting of particles belonging to gate B are being performed from a certain biological sample. In the figure, the sorting operation is being performed so that different sorted products are formed in each of two containers (Right and Left). Sort Gate indicates the gate of the particles to be sorted into each container. Elapsed Time indicates the elapsed time of the sorting operation. Remaining Time indicates the remaining time of the sorting operation. Sort Count indicates the number of particles to be sorted that have been sorted. Sort Rate indicates the rate at which the sorting operation is performed, and eps (events / second) indicates the frequency of the corresponding event. Sort Efficiency indicates the efficiency of sorting. Abort Count indicates the number of aborts.

[0081] Depending on the values ​​related to the sorting operation results, the bioparticle sorting device may display a screen prompting the user to select a sorting mode. For example, as shown in FIG. 5B, a screen 400 may be displayed prompting the user to select a sorting mode based on the Abort Count, particularly the Sort Count and Abort Count. An example of this screen is shown in FIG. 5C. As shown in the figure, the screen displays a message indicating that the Abort Count occurs frequently. Furthermore, the screen displays a message indicating that a high purity and yield of sorted product may be obtained by adopting a different sorting method (Ignore mode). This display can prompt the user to change the sorting mode. Furthermore, the screen displays a button 401 (Sort Settings) for starting to change the sorting mode settings. In response to selecting this button, the bioparticle sorting device may display the sorting operation setting screen as shown in FIG. 4A described above. This allows the sorting mode settings to be easily changed. The screen also displays a button 402 (Close) for closing screen 400. When this button is selected, the bioparticle sorting device closes screen 400. For example, it is conceivable that the user may not want to change the sorting settings in the middle of a sorting operation, and this button can be used to accommodate such cases.

[0082] The bioparticle sorting device may display a screen such as that shown in FIG. 5C that prompts the user to select a sorting mode based on the status of the sorting operation. For example, the bioparticle sorting device may display this screen when the number of Abort Counts is equal to or greater than a predetermined value. The predetermined value may be a value pre-stored in the device or may be a value set by the user. The predetermined value may be set appropriately by a person skilled in the art. In some embodiments, the bioparticle sorting device may display a screen that prompts the user to select a sorting mode depending on the ratio between the Sort Count and the Abort Count. For example, the bioparticle sorting device may display this screen when the Abort Count is, for example, 1 time or more, 1.5 times or more, 2 times or more, 2.5 times or more, or 3 times or more the Sort Count. In this way, by prompting the user to change the sorting mode when the number of Abort Counts is large, a sorting process appropriate for the sample composition can be performed. The Abort Count may refer to the number of times that a particle to be determined is a particle to be sorted, but that it is determined not to sort the particle to be determined due to the presence of other particles present within a predetermined range around the particle to be determined (for example, because the other particles are particles not to be sorted). The Abort Count may be represented by the number of particles belonging to the particle population (b). The Sort Count may refer to the number of times that a particle to be determined is a particle to be sorted and that it is determined to sort the particle to be determined. The Sort Count may be represented by the number of particles belonging to the particle population (a). Therefore, the biological particle sorting device may display a screen on a display unit that prompts the user to select a sorting mode for obtaining the two or more sorted products when the number of particles belonging to the particle population (b) is equal to or greater than a predetermined value, or when the ratio of the number of particles belonging to the particle population (b) to the number of particles belonging to the particle population (a) is equal to or greater than a predetermined value.

[0083] 2. Second embodiment (bioparticle sorting method)

[0084] The present disclosure also relates to a biological particle sorting method. The biological particle sorting method may include: performing a particle sorting determination for particles contained in a biological sample so that particles to be sorted and particles that are not to be sorted but are permissible for sorting are sorted into a first storage space; and displaying information regarding the purity of the particles to be sorted contained in the first storage space. The sorting determination may be performed as performed by the biological particle sorting device (particularly the determination unit) according to the present disclosure described in 1. above, and the same description also applies to the method. The display of the information may be performed as performed by the biological particle sorting device (particularly the display unit) according to the present disclosure described in 1. above, and the same description also applies to the method. The biological particle sorting method may also include performing a particle sorting operation to obtain two or more sorted products from a single biological sample, and the particle sorting operation may be performed so that at least two sorted products having different purities of particles to be sorted are obtained. The biological particle sorting method may include performing a particle sorting operation to obtain two or more sorted products having different purities of particles to be sorted. The biological particle sorting method may include performing a particle sorting operation to obtain two or more sorted products having different purities of particles to be sorted. The biological particle sorting method may include performing a particle sorting operation to obtain two or more sorted products having different purities of particles to be sorted. The method according to the present disclosure may include a particle sorting operation performed by the biological particle sorting device according to the present disclosure described in 1. above, and the description thereof also applies to the method. That is, the biological particle sorting method may be a particle sorting operation for obtaining two or more sorted products from one biological sample, and in the particle sorting operation, at least two sorted products having different purities of particles to be sorted may be obtained. Details of each step performed in the method according to the present disclosure are as described in 1. above, particularly as described with reference to the flow chart of Figure 3, and the description thereof also applies to this embodiment.

[0085] The present disclosure also provides a program for causing a bioparticle sorting device to execute the bioparticle sorting method according to the present disclosure. The program may be stored in, for example, the bioparticle sorting device (e.g., a memory unit) or in an information storage medium. The information storage medium may be, for example, an SD card, a microSD card, a CD, a DVD, a flash memory, or a magnetic recording medium.

[0086] The present disclosure may be configured as follows: [1] A biological particle sorting device comprising: a determination unit that performs particle sorting determination for particles contained in a biological sample so that particles that are target for sorting and particles that are not target for sorting but are acceptable for sorting are sorted into a first storage space; and a display unit that displays information on the purity of the particles that are target for sorting contained in the first storage space. [2] The biological particle sorting device according to [1], wherein the determination unit performs the sorting determination in a sorting mode in which particles contained in the biological sample are sorted into two or more storage spaces including the first storage space and a second storage space, and the sorting mode is configured to perform at least one of the following sorting operations (A) and (B): (A) a sorting operation in which only particles that are target for sorting are sorted into the second storage space; and (B) a sorting operation in which only particles that are target for sorting and particles that are not target for sorting but are acceptable for sorting are sorted into the first storage space. [3] The biological particle sorting device according to [2], wherein the display unit displays information regarding the purity of the particles to be sorted contained in the second storage space. [4] The biological particle sorting device according to any one of [1] to [3], which is configured to be able to display, as information regarding the purity of the particles to be sorted contained in the first storage space, at least one of the following: a ratio of the number of particles belonging to particle group (a) to the total number of particles belonging to particle group (a) in the first storage space; a ratio of the number of particles belonging to particle group (a) to the total number of particles belonging to particle group (a) and (b) in the first storage space; and a ratio of the number of particles belonging to particle group (a) to the total number of particles belonging to particle group (a) and (c) in the first storage space. (a) A particle group of particles to be sorted; (b) A particle group of particles that are not to be sorted and for which sorting is permitted; and (c) A particle group of particles that are not to be sorted and for which sorting is not permitted.[5] The biological particle sorting device according to [3] or [4], which is configured to be able to display, as information regarding the purity of the particles to be sorted contained in the second storage space, at least one of the following: a ratio of the number of particles belonging to particle group (a) to the total number of particles belonging to particle group (a) below in the second storage space; a ratio of the number of particles belonging to particle group (a) to the total number of particles belonging to particle group (a) below in the second storage space; and a ratio of the number of particles belonging to particle group (a) to the total number of particles belonging to particle group (a) below in the second storage space. (a) A particle group of particles to be sorted; (b) A particle group of particles that are not to be sorted but for which sorting is permitted; and (c) A particle group of particles that are not to be sorted and for which sorting is not permitted. [6] The biological particle sorting device according to any one of [2] to [5], wherein the determination unit performs the sorting determination in a sorting mode in which particles contained in the biological sample are sorted into two or more storage spaces including at least a first storage space and a second storage space, and the sorting mode is configured to be able to perform at least one of the following sorting operations (A) and (B): (A) a sorting operation in which only particles to be sorted are sorted into the second storage space; and (B) a sorting operation in which only particles to be sorted and particles that are not to be sorted and are permissible for sorting are sorted into the first storage space, and wherein the purity of the particles to be sorted contained in the first storage space and the purity of the particles to be sorted contained in the second storage space are different from each other.[7] The biological particle sorting device according to any one of [1] to [6], wherein the determination unit is configured to perform the sorting determination in a sorting mode in which particles contained in the biological sample are sorted into two or more storage spaces including the first storage space, and to display, as information regarding the purity of the particles to be sorted, at least one of the following: a ratio of the number of particles belonging to particle group (a) to the total number of particles belonging to particle group (a) below in at least one storage space among the two or more storage spaces; a ratio of the number of particles belonging to particle group (a) to the total number of particles belonging to particle group (a) below in the storage space; and a ratio of the number of particles belonging to particle group (a) to the total number of particles belonging to particle group (a) below in the storage space. (a) A particle group of particles to be sorted; (b) A particle group of particles not to be sorted but for which sorting is permitted; and (c) A particle group of particles not to be sorted and for which sorting is not permitted. [8] The bioparticle sorting device is configured to be able to perform the following sorting operations (A') and (B') in the particle sorting operation in the sorting mode: (A') a sorting operation in which only particles to be sorted are sorted into one storage space; and (B') a sorting operation in which only particles to be sorted and particles that are not to be sorted but are permissible for sorting are sorted into another storage space, wherein the sorted product obtained in the one storage space by the sorting operation (A') and the sorted product obtained in the other storage space by the sorting operation (B') constitute two types of sorted products having different purities.[9] In the particle sorting operation in the sorting mode, the determination unit is configured to determine to which of the following particle populations (a) to (c) a particle to be determined and particles that may be present within a predetermined range around the particle to be determined belong: (a) a particle population of particles to be sorted; (b) a particle population of particles that are not to be sorted and for which sorting is permitted; and (c) a particle population of particles that are not to be sorted and for which sorting is not permitted, and the biological particle sorting device sorts the particle to be determined into the second storage space when the particle to be determined belongs to the particle population (a) and no particles belonging to the particle population (b) or (c) exist within the predetermined range, and sorts the particle to be determined into the first storage space when the particle to be determined belongs to the particle population (a) and particles belonging to the particle population (b) exist within the predetermined range but no particles belonging to the particle population (c). The biological particle sorting device according to any one of [2] to [8].

[10] The biological particle sorting device according to any one of [2] to [9], which is configured to be able to perform particle sorting operations in other sorting modes in addition to the sorting mode.

[11] The biological particle sorting device according to any one of [2] to

[10] , which is configured to display a screen on a display unit prompting a user to select a sorting mode, and which screen is configured to allow the user to select the sorting mode.

[12] The biological particle sorting device according to any one of [2] to

[11] , which performs particle sorting operations in the sorting mode when the number of particles belonging to the particle population (b) is equal to or greater than a predetermined value, or when the ratio of the number of particles belonging to the particle population (b) to the number of particles belonging to the particle population (a) is equal to or greater than a predetermined value.

[13] The bioparticle sorting device according to any one of [2] to

[12] , which is configured to be able to switch sorting modes during a particle sorting operation, and which displays a screen on a display unit prompting the user to select the sorting mode when the number of particles belonging to the particle population (b) is equal to or greater than a predetermined value, or when the ratio of the number of particles belonging to the particle population (b) to the number of particles belonging to the particle population (a) is equal to or greater than a predetermined value.

[14] The bioparticle sorting device according to any one of [1] to

[13] , which is configured to count the number of particles that are not to be sorted but are permissible for sorting.

[15] The bioparticle sorting device according to any one of [1] to

[14] , which is configured as a flow cytometer that sorts bioparticles in an open space or a closed space.

[16] The bioparticle sorting device according to any one of [1] to

[15] , wherein the bioparticle sorting device is an imaging flow cytometer.

[17] A bioparticle sorting method comprising: executing particle sorting determination for particles contained in a biological sample so that particles to be sorted and particles that are not to be sorted but are acceptable for sorting are sorted into a first storage space; and displaying information regarding the purity of the particles to be sorted contained in the first storage space.

[0087] 100 Biological particle sorting device 101 Light irradiation unit 102 Detection unit 103 Information processing unit 6100 Biological sample analyzer (biological particle sorting device) 6101 Light irradiation unit 6102 Detection unit 6103 Information processing unit

Claims

1. A biological particle sorting device having a judgment unit that performs particle sorting judgment for particles contained in a biological sample so that particles to be sorted and particles that are not to be sorted but are acceptable for sorting are sorted into a first storage space, and a display unit that displays information regarding the purity of the particles to be sorted that are contained in the first storage space.

2. The biological particle sorting device according to claim 1, wherein the determination unit performs the sorting determination in a sorting mode in which particles contained in the biological sample are sorted into two or more storage spaces including the first storage space and the second storage space, and the sorting mode is configured to perform at least one of the following sorting operations (A) and (B): (A) a sorting operation in which only particles to be sorted are sorted into the second storage space; and (B) a sorting operation in which only particles to be sorted and particles that are not to be sorted and are permissible for sorting are sorted into the first storage space.

3. The bioparticle sorting device according to claim 2, wherein the display unit displays information regarding the purity of the particles to be sorted contained in the second storage space.

4. The biological particle sorting device according to claim 1, which is configured to be able to display, as information regarding the purity of the particles to be sorted contained in the first storage space, at least one of the following: a ratio of the number of particles belonging to particle group (a) to the total number of particles belonging to particle groups (a) to (c) below in the first storage space; a ratio of the number of particles belonging to particle group (a) to the total number of particles belonging to particle groups (a) and (b) below in the first storage space; and a ratio of the number of particles belonging to particle group (a) to the total number of particles belonging to particle groups (a) and (c) below in the first storage space. (a) A particle group of particles to be sorted; (b) A particle group of particles that are not to be sorted and for which sorting is permitted; and (c) A particle group of particles that are not to be sorted and for which sorting is not permitted.

5. The biological particle sorting device according to claim 3, which is configured to be able to display, as information regarding the purity of the particles to be sorted contained in the second storage space, at least one of the following: a ratio of the number of particles belonging to particle group (a) to the total number of particles belonging to particle group (a) below (a) to (c) below in the second storage space; a ratio of the number of particles belonging to particle group (a) to the total number of particles belonging to particle group (a) below (b) below in the second storage space; and a ratio of the number of particles belonging to particle group (a) to the total number of particles belonging to particle group (a) below (c) below in the second storage space. (a) A particle group of particles to be sorted; (b) A particle group of particles not to be sorted and for which sorting is permitted; and (c) A particle group of particles not to be sorted and for which sorting is not permitted.

6. The biological particle sorting device of claim 2, wherein the judgment unit performs the sorting judgment in a sorting mode in which particles contained in the biological sample are sorted into two or more storage spaces including at least a first storage space and a second storage space, and the sorting mode is configured to perform at least one of the following sorting operations (A) and (B): (A) a sorting operation in which only particles to be sorted are sorted into the second storage space; and (B) a sorting operation in which only particles to be sorted and particles that are not to be sorted and are acceptable for sorting are sorted into the first storage space, and the purity of the particles to be sorted contained in the first storage space and the purity of the particles to be sorted contained in the second storage space are different from each other.

7. The biological particle sorting device according to claim 1, wherein the judgment unit is configured to execute the sorting judgment in a sorting mode in which particles contained in the biological sample are sorted into two or more storage spaces including the first storage space, and to display at least one of the following information regarding the purity of the particles to be sorted: a ratio of the number of particles belonging to particle group (a) to the total number of particles belonging to particle groups (a) to (c) in at least one of the two or more storage spaces; a ratio of the number of particles belonging to particle group (a) to the total number of particles belonging to particle groups (a) and (b) in the storage space; and a ratio of the number of particles belonging to particle group (a) to the total number of particles belonging to particle groups (a) and (c) in the storage space. (a) A particle group of particles to be sorted; (b) A particle group of particles that are not to be sorted and for which sorting is permitted; and (c) A particle group of particles that are not to be sorted and for which sorting is not permitted.

8. The bioparticle sorting device is configured to be able to perform the following sorting operations (A') and (B') in a particle sorting operation in the sorting mode: (A') a sorting operation of sorting only particles to be sorted into one storage space; and (B') a sorting operation of sorting only particles to be sorted and particles that are not to be sorted and for which sorting is permitted into another storage space. The bioparticle sorting device described in claim 7, wherein the sorting product obtained in the one storage space by the sorting operation (A') and the sorting product obtained in the other storage space by the sorting operation (B') constitute two types of sorting products having different purities.

9. The biological particle sorting device according to claim 2, wherein in a particle sorting operation in the sorting mode, the determination unit is configured to determine to which of the following particle populations (a) to (c) a particle to be determined and particles that may be present within a predetermined range around the particle to be determined belong: (a) a particle population of particles to be sorted; (b) a particle population of particles not to be sorted and for which sorting is permitted; and (c) a particle population of particles not to be sorted and for which sorting is not permitted; and wherein the biological particle sorting device sorts the particle to be determined into the second storage space when the particle to be determined belongs to the particle population (a) and no particles belonging to the particle population (b) or (c) are present within the predetermined range, and sorts the particle to be determined into the first storage space when the particle to be determined belongs to the particle population (a) and there are particles belonging to the particle population (b) but no particles belonging to the particle population (c) within the predetermined range.

10. The bioparticle sorting device according to claim 2, wherein the bioparticle sorting device is configured to be able to perform particle sorting operations in other sorting modes in addition to the sorting mode.

11. The bioparticle sorting device of claim 2, wherein the bioparticle sorting device is configured to display a screen on a display unit that prompts a user to select a sorting mode, and the screen is configured to allow the user to select the sorting mode.

12. The bioparticle sorting device according to claim 2, wherein the bioparticle sorting device performs a particle sorting operation in the sorting mode when the number of particles belonging to the particle population (b) is equal to or greater than a predetermined value, or when the ratio of the number of particles belonging to the particle population (b) to the number of particles belonging to the particle population (a) is equal to or greater than a predetermined value.

13. The bioparticle sorting device of claim 2, wherein the sorting mode can be switched during a particle sorting operation, and the bioparticle sorting device displays a screen on a display unit that prompts the user to select the sorting mode when the number of particles belonging to the particle population (b) is equal to or greater than a predetermined value, or when the ratio of the number of particles belonging to the particle population (b) to the number of particles belonging to the particle population (a) is equal to or greater than a predetermined value.

14. The bioparticle sorting device according to claim 1, wherein the bioparticle sorting device is configured to count the number of particles that are not subject to sorting and are permissible for sorting.

15. The bioparticle sorting device according to claim 1, which is configured as a flow cytometer that sorts bioparticles in an open space or in a closed space.

16. The bioparticle sorting device according to claim 1, wherein the bioparticle sorting device is an imaging flow cytometer.

17. A biological particle sorting method comprising: executing a particle sorting determination for particles contained in a biological sample so as to sort particles to be sorted and particles that are not to be sorted but are acceptable for sorting into a first storage space; and displaying information regarding the purity of the particles to be sorted contained in the first storage space.

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