Particle analysis system, information processing method, and program

Abstract

The main purpose of the present technology is to improve the ability to separate a fluorescence spectrum.　The present technology provides a particle analysis system including: a light irradiation unit that includes at least one first light source that emits light having a wavelength of not less than 350 nm and at least one second light source that emits light having a wavelength of less than 350 nm; and a processing unit that performs an unmixing process on optical data obtained by irradiation of particles with light by the light irradiation unit. In addition, the present technology provides an information processing method including an unmixing process step for performing an unmixing process on optical data obtained by irradiation of particles with light by a light irradiation unit that includes at least one first light source emitting light having a wavelength of not less than 350 nm and at least one second light source emitting light having a wavelength of less than 350 nm.

Description

Technical Field

[0001] The present technology relates to a particle analysis system, an information processing method, and a program. More specifically, the present technology relates to a particle analysis system that analyzes particles based on light generated by irradiating particles with light, and an information processing method and a program used in the particle analysis system.

Background Art

[0002] For example, a particle population such as cells is labeled with a fluorescent dye, and the characteristics of the particles are measured by irradiating each particle of the particle population with laser light and measuring the intensity and / or pattern of fluorescence generated from the excited fluorescent dye. As a typical example of a particle analyzer for performing such measurement, a flow cytometer can be mentioned.

[0003] A fluorescent dye is associated with an excitation wavelength at which a signal can be obtained at a high peak. Therefore, in a conventional flow cytometer, particles labeled with a fluorescent dye are irradiated with light having the excitation wavelength thereof, and a fluorescence signal is obtained using an optical filter corresponding to the peak wavelength range of the fluorescent dye.

[0004] On the other hand, in a spectral flow cytometer, for example, the fluorescence of each cell is collectively obtained as a spectrum. Fluorescence separation processing (also called unmixing processing) is performed on the obtained fluorescence spectrum using a spectral reference of each fluorescent dye, and a fluorescence signal is obtained.

[0005] As a technique related to the fluorescence separation processing, for example, in Patent Document 1 below, fluorescence generated from a fluorescent dye excited by irradiating microparticles multiply labeled with a plurality of fluorescent dyes having overlapping fluorescence wavelength bands with light is received by photodetectors having different received wavelength bands arranged in a number larger than the number of fluorescent dyes, and a measurement spectrum obtained by collecting detection values from each photodetector is approximated by a linear sum of single-staining spectra obtained with microparticles individually labeled with each fluorescent dye. A fluorescence intensity correction method including the procedure is disclosed.

Prior Art Documents

Patent Documents

[0006]

Patent Document 1

Summary of the Invention

Problems to be Solved by the Invention

[0007] For example, in the development of cancer immunotherapy, a flow cytometer capable of analyzing multi-color fluorescent dyes is often used to elucidate the immune mechanism. In a spectral type flow cytometer, as described above, by separating the fluorescence spectrum acquired all at once by unmixing processing, it is possible to analyze more particles labeled with fluorescent dyes than a conventional type flow cytometer. However, for example, in order to elucidate a complex immune mechanism, more improved separation performance may be required.

[0008] Therefore, the present technology mainly aims to improve the separation performance of fluorescence spectra.

Means for Solving the Problems

[0009] The present technology provides an optical irradiation unit including at least one first light source that emits light having a wavelength of 350 nm or more and at least one second light source that emits light having a wavelength of less than 350 nm, and a processing unit that performs unmixing processing on the optical data obtained by irradiating particles with the light from the optical irradiation unit. A particle analysis system including the above is provided. The at least one second light source can emit light having a wavelength of 250 nm or more and less than 350 nm. The particle analysis system may be configured such that at least two excitation lights out of the excitation light emitted by the at least one first light source and the excitation light emitted by the at least one second light source are combined, and the combined excitation light irradiates particles. The particle analysis system may be configured to analyze a population of particles labeled with a plurality of phosphors. The processing unit can execute the unmixing process using spectral reference data. In the particle analysis system, spectral data of fluorescence regarding a population of particles labeled with each of the plurality of phosphors may be used as the spectral reference data used in the unmixing process. The particle analysis system may further include a detection unit that detects light generated by irradiating the particles with light from the light irradiation unit. The detection unit may include at least one photodetector that detects light generated by irradiating the particles with light from the light irradiation unit. The at least one photodetector has a light receiving element array, and The processing unit can acquire only a signal based on light received by a part of the light receiving elements constituting the light receiving element array according to the wavelength of the light. The at least one photodetector may be configured such that a part of the light receiving elements constituting the light receiving element array does not perform signal transmission according to the wavelength of the light. The at least one photodetector may be controlled such that a part of the light receiving elements constituting the light receiving element array does not perform signal transmission. In a preferred embodiment of the present technology, the detection unit may include a plurality of photodetectors. Each of the plurality of photodetectors is associated with a light source included in the light irradiation unit. The processing unit can acquire only a signal based on light received by a part of the light receiving elements constituting the light receiving element array of each photodetector according to the wavelength of the light source associated with each photodetector. In the preferred embodiment, each of the plurality of photodetectors is configured not to transmit a signal of light having a wavelength equal to or less than the wavelength of the associated light source, and Among the light-receiving elements constituting the light-receiving element array of each photodetector, only the light-receiving elements that receive light having a wavelength longer than the wavelength of the associated light source may be connected to a signal transmission circuit that transmits a signal based on the received light. In the preferred embodiment, each of the plurality of photodetectors is controlled so as not to transmit a signal of light having a wavelength equal to or less than the wavelength of the associated light source, and each photodetector can be controlled such that, among the light-receiving elements constituting the light-receiving element array, only the light-receiving elements that receive light having a wavelength longer than the wavelength of the associated light source transmit a signal based on the received light. The plurality of photodetectors may have the same light-receiving element array. The at least one first light source may be a laser light source, and the at least one second light source may also be a laser light source.

[0010] The present technology also provides an information processing method including an unmixing process for unmixing light data obtained by irradiating particles with light from a light irradiation unit including at least one first light source that emits light having a wavelength of 350 nm or more and at least one second light source that emits light having a wavelength of less than 350 nm.

[0011] The present technology also provides a program for causing an information processing apparatus to execute an unmixing process for unmixing light data obtained by irradiating particles with light from a light irradiation unit including at least one first light source that emits light having a wavelength of 350 nm or more and at least one second light source that emits light having a wavelength of less than 350 nm.

Brief Description of the Drawings

[0012]

Figure 1

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Embodiments for Carrying Out the Invention

[0013] Hereinafter, preferred embodiments for carrying out the present technology will be described. Note that the embodiments described below show typical embodiments of the present technology, and the scope of the present technology is not limited only to these embodiments. The description of the present technology will be made in the following order. 1. First Embodiment (Particle Analysis System) (1) Description of the First Embodiment (2) First Example of the First Embodiment (2-1) Light Irradiation Unit (2-2) Chip (2-3) Detection Unit (2-3-1) Example of the Detection Unit (2-3-2) Example of the Detection Unit (2-4) Example of the Configuration of the Optical System (2-5) Information Processing Device (2-6) Output Unit and Input Unit (2-7) Example of Information Processing by the Information Processing Device (2-8) Particles 2. Second Embodiment (Particle Analyzer) 3. Third Embodiment (Information Processing Method) 4. Fourth Embodiment (Program) 5. Examples

[0014] 1. First Embodiment (Particle Analysis System)

[0015] (1) Description of the First Embodiment

[0016] The particle analysis system according to the present technology includes an optical irradiation unit including at least one first light source that emits light with a wavelength of 350 nm or more and at least one second light source that emits light with a wavelength of less than 350 nm. By combining and using the first light source that emits light on the long wavelength side and the second light source that emits light on the short wavelength side as described above, the fluorescence separation performance in the unmixing process can be improved.

[0017] The present technology is suitable when the number of types of phosphors used is large, such as in multi-color analysis. For example, even when there are 10 or more types, particularly 15 or more types, more particularly 20 or more types, further 25 or more types, 30 or more types, 35 or more types, or 40 or more types, by applying the present technology, more appropriate fluorescence separation processing becomes possible. The present technology may be applied for the analysis of a particle population labeled with such a number of types of phosphors.

[0018] (2) First Example of the First Embodiment

[0019] The particle analysis system according to the present technology may be configured as a particle analysis system that performs, for example, flow cytometry. An example of the particle analysis system according to the present technology configured in this way and an example of the processing by the particle analysis system will be described below with reference to FIG. 1. FIG. 1 shows a configuration example of the particle analysis system according to the present technology.

[0020] The particle analysis system 1 shown in Fig. 1 includes a light irradiation unit 2, a chip T provided with a flow path through which particles to be analyzed flow, a detection unit 3, an information processing device 100, an output unit 4, and an input unit 5. The particle analysis system 1 is configured as a system for performing flow cytometry. For example, the light irradiation unit 2 and the detection unit 3 may be configured as one particle analysis device, and the particle analysis device may be combined with the information processing device 100 to be configured as a particle analysis system. The particle analysis device may be connected to the information processing device 100 by wire or wirelessly, or may be connected via a network. The output unit 4 and the input unit 5 may be provided in the particle analysis device or the information processing device 100, or may be configured as a device separate from the particle analysis device and the information processing device 100.

[0021] (2-1) Light irradiation unit

[0022] The light irradiation unit 2 is configured to irradiate light at a predetermined position in the flow path of the chip T. When particles pass through the light irradiation position in the flow path, the particles are irradiated with light, and as a result, fluorescence is generated. That is, the light can act as excitation light on the particles, particularly on the phosphor that labels the particles.

[0023] The light irradiation unit 2 includes at least one first light source that emits light with a wavelength of 350 nm or more and at least one second light source that emits light with a wavelength of less than 350 nm. By combining the first light source and the second light source, for example, compared with the case of using only the first light source, the fluorescence separation performance in the case of unmixing treatment can be improved. The at least one first light source may be a laser light source, but may also be other light sources, such as an LED. The at least one second light source may also be a laser light source, but may also be other light sources, such as an LED. Preferably, the at least one first light source is a laser light source, and the at least one second light source is a laser light source.

[0024] The at least one first light source preferably emits light with a wavelength of 900 nm or less, more preferably 880 nm or less, and even more preferably 850 nm or less. For example, the at least one first light source may be a light source that emits light with a wavelength of 350 nm or more and 900 nm or less, more preferably 350 nm or more and 880 nm or less, and even more preferably 350 nm or more and 850 nm or less. The light is, for example, laser light, that is, the at least one first light source may be a laser light source, which also applies to the following description of the first light source. The number of the at least one first light source can be, for example, 1 to 20, particularly 2 to 15, and more particularly 3 to 10.

[0025] The at least one second light source is preferably a light source that emits light with a wavelength of 250 nm or more, more preferably 260 nm or more, even more preferably 270 nm or more, and particularly preferably 280 nm or more. By setting the wavelength of the light of the second light source to the above lower limit value or more, autofluorescence generated by light irradiation of particles (particularly cells) can be reduced, which contributes to the improvement of separation performance. Preferably, the at least one second light source may be a light source that emits light with a wavelength of 250 nm or more and less than 350 nm, preferably 260 nm or more and less than 350 nm, more preferably 270 nm or more and less than 350 nm, and even more preferably 280 nm or more and less than 350 nm. The light is, for example, laser light, that is, the at least one second light source may be a laser light source, which also applies to the following description of the second light source. The number of the at least one second light source can be, for example, 1 to 10, particularly 1 to 5, and more particularly 1 to 3.

[0026] According to one embodiment of the present technology, the at least one second light source may include at least one (for example, 1, 2, or 3) light source that emits light with a wavelength of 305 nm or more and less than 350 nm, 310 nm or more and less than 350 nm, or 315 nm or more and less than 350 nm. According to another embodiment of the present technology, the at least one second light source may include at least one (e.g., 1, 2, or 3) light source that emits light having a wavelength of less than 305 nm, less than 310 nm, or less than 315 nm. According to still another embodiment of the present technology, the at least one second light source may include at least one (e.g., 1, 2, or 3) light source that emits light having a wavelength of less than 305 nm, less than 310 nm, or less than 315 nm, and at least one (e.g., 1, 2, or 3) light source that emits light having a wavelength of 305 nm or more and less than 350 nm, 310 nm or more and less than 350 nm, or 315 nm or more and less than 350 nm.

[0027] According to one embodiment of the present technology, the wavelength of the at least one second light source may be, for example, 20 nm or more, 30 nm or more, or 40 nm or more smaller than the minimum wavelength among the wavelengths of the at least one first light source.

[0028] The wavelength of the at least one second light source may preferably be different from any of the excitation peak wavelengths of the phosphor that labels the particle population to be analyzed by the particle analysis system of the present technology. The wavelength of the at least one second light source is, for example, preferably 5 nm or more smaller than the excitation peak wavelength of the phosphor that labels the particle population to be analyzed and has the excitation peak wavelength closest to the wavelength of the second light source (or the phosphor having the smallest excitation peak wavelength), more preferably 10 nm or more smaller, even more preferably 15 nm or more, 20 nm or more, or 25 nm or more smaller. In this way, using the light obtained by irradiating the particles with light having a wavelength not associated with the excitation peak wavelength of the phosphor contributes to improving the fluorescence separation performance.

[0029] The total number of the first light source and the second light source is 2 or more, and may be, for example, 3 or more, particularly 4 or more, and more particularly 5 or more. The total number may be, for example, 30 or less, particularly 20 or less, and more particularly 15 or less.

[0030] Each of the at least one first light source and the at least one second light source may be a laser light source that emits laser light of a single wavelength, for example, a laser light source with a fixed oscillation wavelength or a laser light source with a variable oscillation wavelength. The wavelengths of these laser light sources mean the oscillation wavelengths. The laser light emitted by these laser light sources may irradiate the particles as it is at its oscillation wavelength.

[0031] When the first light source is a laser light source, the first light source may be any one selected from the group consisting of a semiconductor laser, an argon ion (Ar) laser, a helium-neon (He-Ne) laser, a dye laser, a krypton (Cr) laser, and a solid-state laser combined with a wavelength conversion optical element, and particularly preferably a semiconductor laser. Alternatively, the first light source may be an LED. The second light source may also be any laser light source selected from the group mentioned for the first light source, and particularly preferably a semiconductor laser. Alternatively, the second light source may be an LED.

[0032] The light irradiation unit 2 may be configured such that at least two of the excitation lights (particularly laser lights) emitted by the at least one first light source and the excitation lights (particularly laser lights) emitted by the at least one second light source are combined, and the combined excitation light irradiates the particles. That is, the light irradiation unit 2 may be configured such that a plurality of excitation lights are combined and irradiated onto one or more (for example, 1, 2, 3, 4, or 5) spots, and the particle analysis system 1 may be configured such that the particles pass through the spots. To configure the light irradiation unit 2 in this way, the light irradiation unit 2 may include a light guiding optical system for guiding these plurality of excitation lights to a predetermined position. The light guiding optical system may include optical components such as a group of beam splitters and a group of mirrors, for example, to combine the plurality of excitation lights. Further, the light guiding optical system may include a group of lenses for condensing the combined excitation light, and may include an objective lens, for example.

[0033] (2-2) Chip

[0034] Chip T can be configured as a flow cell, for example. A flow path is provided in chip T. The flow path structure provided in chip T is configured to form a flow (particularly a laminar flow) in which particles flow in a substantially straight line, for example.

[0035] In chip T shown in FIG. 1, flow paths P11, P12a, P12b, and P13 are provided. A sample liquid containing particles is introduced from a container (bag) B1 containing the sample liquid into the sample liquid flow path P11. The sample liquid flows through the sample liquid flow path P11 toward the main flow path P13. A sheath liquid is introduced into chip T from a container (bag) B2 containing the sheath liquid. The sheath liquid flows through the two sheath liquid flow paths P12a and P12b toward the main flow path P13. The sample liquid flow path P11 and the sheath liquid flow paths P12a and P12b are configured to merge to form the main flow path P13. The sample liquid fed in the sample liquid flow path P11 and the sheath liquid fed in the sheath liquid flow paths P12a and P12b merge at the point where the three flow paths merge and then flow in the main flow path P13. In the main flow path P13, for example, a laminar flow in which the sample liquid is sandwiched between the sheath liquids flows. In the laminar flow, the particles are arranged in a substantially straight line. The particles flowing side by side in the main flow path P13 are irradiated with light (particularly laser light) generated by the light irradiation unit 2, and the light generated thereby is detected by the detection unit 3.

[0036] Chip T may have a two-dimensional or three-dimensional flow path structure. Chip T may have a substrate shape formed from a plastic material or a glass material. Chip T and the flow path structure provided in chip T are not limited to those shown in FIG. 1, and for example, chips and flow path structures known in the technical field related to flow cytometers may be adopted. That is, in the present technology, fluorescence detection may be fluorescence detection by a flow cytometer, for example. The cross-sectional shape of the flow path provided in the chip T may be, for example, circular, elliptical, or rectangular (square or rectangular), etc. When the cross-section of the flow path is circular or elliptical, its diameter or major axis may be, for example, 1 mm or less, and particularly may be 10 μm or more and 1 mm or less. When the cross-section of the flow path is square or rectangular, the length of one side or the long side may be, for example, 1 mm or less, and particularly may be 10 μm or more and 1 mm or less.

[0037] The particles emitted from the chip T may be sorted. For example, by vibrating the chip with a vibrating element such as a piezoelectric vibrating element, droplets containing one particle can be generated from the discharge port. By charging the droplets with a charging unit, the traveling direction can be controlled and the particles can be sorted. Thus, the particle analysis system 1 may be configured as a system having a sorting function. Also, as the chip T, a chip provided with a sorting mechanism inside the chip may be used. As an example of such a chip, for example, the microchip for microparticle sorting described in JP-A-2019-174192 can be cited. With this chip, particles in the sample liquid can be sorted without contact with the outside air, that is, a closed-type sorting operation is possible. As described above, the particle analysis system 1 may have a sorting unit for sorting particles. The sorting unit can sort particles based on the fluorescence detection result by the detection unit.

[0038] (2-3) Detection unit

[0039] The detection unit 3 detects the light generated by the light irradiation of the particles by the light irradiation unit 2. For example, the detection unit 3 may be configured to detect the light generated by the light irradiation of the particles flowing in the flow path of the chip T. The light detected by the detection unit 3 is, for example, light containing fluorescence, and may be light containing both fluorescence and light other than fluorescence. The detection unit 3 may be further configured to detect scattered light (for example, any one or more of forward scattered light, backward scattered light, and lateral scattered light) in addition to the detection of fluorescence.

[0040] The detection unit 3 includes at least one photodetector that detects the light generated by irradiating the particles with the light irradiation unit 2. The number of photodetectors provided in the detection unit 3 may be, for example, 1 to 20, 1 to 15, or 1 to 10. Each photodetector includes one or more light-receiving elements and, for example, has a light-receiving element array. Each photodetector may include, for example, as the light-receiving element, one or more PMTs (photomultiplier tubes) and / or photodiodes, and particularly includes one or more PMTs. The photodetector may include, for example, a PMT array in which a plurality of PMTs are arranged in a one-dimensional direction. The at least one photodetector may be one that detects fluorescence and may be configured as a fluorescence detector. The number of light-receiving elements (for example, the number of PMTs) included in each photodetector may be, for example, 2 or more, 5 or more, 8 or more, 10 or more, 15 or more, 20 or more, 22 or more, 24 or more, or 26 or more. The number of light-receiving elements (for example, the number of PMTs) included in each photodetector may be, for example, 50 or less, 45 or less, or 40 or less.

[0041] The detection unit 3 may include a spectroscopic unit that spectroscopically analyzes light. The spectroscopic unit may be provided in each photodetector. The spectroscopic unit may be configured to, for example, spectroscopically analyze light (for example, fluorescence) and cause the light of a predetermined detection wavelength to reach a light-receiving element (for example, a PMT) to which the predetermined detection wavelength is assigned.

[0042] Each photodetector included in the detection unit 3 includes a transmission unit that transmits the signal received by the light-receiving element. The transmission unit may include a signal transmission circuit connected to the light-receiving element. The signal transmission circuit may be configured as an amplification circuit, for example. The transmission unit amplifies the received signal, for example, and transmits it to a signal processing unit described later.

[0043] When the at least one photodetector provided in the detection unit 3 has a light receiving element array, the processing unit 101 described later can acquire only the signal based on the light received by a part of the light receiving elements constituting the light receiving element array according to the wavelength of the light. In order to enable such signal acquisition by the processing unit 101, in a preferred embodiment of the present technology, the photodetector included in the detection unit 3 may be configured such that a part of the light receiving elements constituting the light receiving element array does not perform signal transmission according to the wavelength of the light (more specifically, according to the wavelengths of the first light source and the second light source), or a part of the light receiving elements constituting the light receiving element array may be controlled so as not to perform signal transmission. Thereby, the amount of signal transmission can be reduced.

[0044] More specific configuration examples of the detection unit 3 and the photodetector included in the detection unit 3 will be described in the following (2-3-1) and (2-3-2). Regarding the configuration of the photodetector that enables the reduction of the signal transmission amount described above, it will be described in the following (2-3-2).

[0045] The detection unit 3 may include one or more measuring instruments selected from a fluorescence measuring instrument, a scattered light measuring instrument, a transmitted light measuring instrument, a reflected light measuring instrument, a diffracted light measuring instrument, an ultraviolet spectroscopic measuring instrument, an infrared spectroscopic measuring instrument, a Raman spectroscopic measuring instrument, a FRET measuring instrument, and a FISH measuring instrument. Further, the detection unit 3 may include a two-dimensional light receiving element such as a CCD or a CMOS, for example.

[0046] The detection unit 3 may include a signal processing unit. The signal processing unit converts the electrical signal obtained by the fluorescence detector into a digital signal. The signal processing unit may include, for example, an A / D converter as a device for performing the conversion. The optical signal detected by the photodetector can be converted into a digital signal by the signal processing unit and then transmitted to the information processing device 100. The digital signal can be handled as optical data by the information processing device 100 and can be the target of the unmixing processing by the processing unit described later. The optical data may include data regarding fluorescence intensity.

[0047] The detection unit 3 (particularly the photodetector) is arranged at a position where it can detect the light generated from the particles. For example, as shown in FIG. 1, the detection unit 3 may be arranged so as to sandwich the chip T (particularly the main flow path P13) between the light irradiation unit 2 and the detection unit 3, or the detection unit 3 may be arranged on the same side of the chip T as the light irradiation unit 2.

[0048] (2-3-1) Example of the detection unit

[0049] An example of the photodetector included in the detection unit 3 will be described below with reference to FIG. 3.

[0050] The photodetector 300 shown in FIG. 3 includes a spectroscopic unit 301, a light receiving element array 302, and a transmission unit 303.

[0051] The spectroscopic unit 301 spectroscopically analyzes the light generated by irradiating the particles with the light from the light irradiation unit 2, and guides the spectroscopically analyzed light to the light receiving element array 302. The spectroscopic unit 301 includes, for example, one or a plurality of prisms. In particular, the spectroscopic unit 301 includes a prism array including a plurality of prisms. The configuration of the spectroscopic unit 301 may be appropriately set according to the configuration of the light receiving element array 302. In FIG. 3, the spectroscopic unit 301 spectroscopically analyzes the light into eight lights with wavelengths from a longer wavelength λ1 to a shorter wavelength λ8. That is, λ1~λ8 are in the relationship of λ1>λ2>λ3>λ4>λ5>λ6>λ7>λ8.

[0052] The light receiving element array 302 receives the light spectroscopically analyzed by the spectroscopic unit 301. As shown in FIG. 3, the light receiving element array 302 may include a plurality of arranged light receiving elements 302-1 to 302-8. The light receiving element 302-1 receives the light with wavelength λ1, and similarly, the light receiving elements 302-2 to 302-8 receive the lights with wavelengths λ2 to λ8, respectively. In FIG. 3, the number of light receiving elements included in one light receiving element array 302 is eight, but the number of light receiving elements is not limited to this. The number of light receiving elements included in the light receiving element array 302 may be, for example, two or more, five or more, eight or more, ten or more, fifteen or more, twenty or more, twenty-two or more, twenty-four or more, or twenty-six or more. The number of light receiving elements included in the light receiving element array 302 may also be, for example, fifty or less, forty-five or less, or forty or less.

[0053] The light receiving elements 302-1 to 302-8 included in the light receiving element array 302 are light sensors and may be, for example, PMTs. That is, the light receiving element array 302 may be a PMT array. The PMT can, for example, sensitively detect weak fluorescence generated by irradiating cells labeled with a fluorescent dye with light. Each light receiving element receives each light split by the splitting unit 301 and converts it into an electrical signal. Each light receiving element transmits the converted electrical signal to a transmission circuit connected to each light receiving element.

[0054] The transmission unit 303 transmits the electrical signal acquired by the light receiving element array 302 to the outside of the photodetector 300, for example, to the information processing device 100 connected to the photodetector 300. The transmission unit 303 includes transmission circuits 303-1 to 303-8 connected to each light receiving element of the light receiving element array 302. As shown in FIG. 3, one transmission circuit may be connected to one light receiving element. Each transmission circuit may be configured as an amplifier circuit that amplifies the electrical signal obtained by the light receiving element. The electrical signal based on light is amplified by the amplifier circuit and transmitted to, for example, a signal processing unit or an information processing device outside the photodetector 300.

[0055] (2-3-2) Example of the detection unit

[0056] In one preferred embodiment of the present technology, the detection unit includes a plurality of photodetectors. In this embodiment, each of the plurality of photodetectors may be associated with any one of the light sources included in the light irradiation unit. More specifically, each of the plurality of photodetectors may be associated with any one of the at least one first light source and the at least one second light source. In this embodiment, the processing unit can acquire only signals based on the light received by a part of the light receiving elements constituting the light receiving element array of each photodetector according to the wavelength of the light source associated with each photodetector. To enable such signal acquisition, for example, each of the plurality of photodetectors may be configured not to transmit signals of light having a wavelength equal to or less than the wavelength of the associated light source, or may be controlled not to transmit signals of light having a wavelength equal to or less than the laser light wavelength of the associated light source. In this specification, "the wavelength of the light source" means the wavelength of the light emitted by the light source. When the light source is a laser light source, "the wavelength of the light source" may mean the wavelength of the laser light emitted by the laser light source. In this embodiment, preferably, the plurality of photodetectors have the same light receiving element array. Thereby, the detection unit can be configured more simply. By configuring the detection unit according to this embodiment, the amount of signals transmitted can be reduced. The reduction of the amount of transmitted signals will be described in more detail below.

[0057] In particle analysis using a plurality of fluorescent dyes such as the multi-color analysis described above, as more phosphors label particles (especially cells), the amount of signals acquired by fluorescence detection increases. The increase in the amount of signals can lead to, for example, an increase in data transmission time, an increase in the burden of data processing, and an increase in the data occupancy ratio in storage. The fluorescence generated from the phosphor has a wavelength longer than that of the excitation light. Therefore, a fluorescence detector assigned to detect the fluorescence generated by a predetermined excitation light does not need to detect light having a wavelength equal to or shorter than the wavelength of the predetermined excitation light. In this embodiment, the photodetector is configured not to transmit a signal of light having a wavelength equal to or shorter than the wavelength of the associated light source, or is controlled not to transmit a signal of light having a wavelength equal to or shorter than the wavelength of the associated light source. This prevents light that does not need to be detected from being transmitted, resulting in a reduction in the amount of signal transmitted. This can lead to a reduction in data transmission time, a reduction in the burden of data processing, and a reduction in the data occupancy ratio in storage.

[0058] For example, one photodetector may be associated with one light source, or two or more photodetectors may be associated with one light source.

[0059] An example in which each of a plurality of photodetectors is configured not to transmit a signal of fluorescence having a wavelength equal to or shorter than the wavelength of the associated light source in this embodiment will be described with reference to FIG. 4. In this example, each photodetector is connected to a signal transmission circuit in which only the light receiving elements that receive light having a wavelength longer than the wavelength of the associated light source among the light receiving element array transmit a signal based on the received light.

[0060] The detection unit 3 shown in FIG. 4 includes three photodetectors 400a, 400b, and 400c.

[0061] The photodetector 400a includes a spectroscopic unit 401a, a light receiving element array 402a, and a transmission unit 403a. The photodetector 400a, and the spectroscopic unit 401a, the light receiving element array 402a, and the transmission unit 403a included therein are the same as the photodetector 300, and the spectroscopic unit 301, the light receiving element array 302, and the transmission unit 303 described in (2-3-1) above, and the description thereof also applies to this example.

[0062] The photodetector 400a is associated with a predetermined light source a (not shown) included in the light irradiation unit 2. The light source a emits light (particularly laser light) having a wavelength λa. The wavelength λa is shorter than any of the wavelengths of the light λ1 to λ8 detected by the light receiving element array 402a. The light having the wavelength λa emitted by the light source a may generate fluorescence having wavelengths λ1 to λ8 that are longer than the wavelength λa. Therefore, transmission circuits are respectively connected to all the light receiving elements.

[0063] The photodetector 400b includes a spectroscopic unit 401b, a light receiving element array 402b, and a transmission unit 403b. The photodetector 400b, and the spectroscopic unit 401b and the light receiving element array 402b included therein are the same as the photodetector 300 described in (2-3-1) above, and the spectroscopic unit 301 and the light receiving element array 302, and the description thereof also applies in this example. On the other hand, the transmission unit 403b is different from the transmission unit 303 described in (2-3-1) above, that is, it is also different from the transmission unit 403a of the photodetector 400a. The difference is that transmission circuits are connected only to some of the light receiving elements included in the light receiving element array 402b, and transmission circuits are not connected to the remaining light receiving elements. More specifically, transmission circuits 403b-1 to 403b-6 are respectively connected to the light receiving elements 402b-1 to 402b-6, while no transmission circuit is connected to the light receiving elements 402b-7 to 402b-8. In this way, the transmission unit 403b is configured not to transmit signals detected by some of the light receiving elements in the light receiving element array 402b.

[0064] The photodetector 400b is associated with a predetermined light source b (not shown) included in the light irradiation unit 2. The light source b emits light (particularly laser light) having a wavelength λb. The wavelength λb is shorter than λ6 and longer than λ7 among the wavelengths λ1 to λ8 of light that can be detected by the light receiving element array 402b. Therefore, the laser light having the wavelength λb emitted by the light source b may generate fluorescence having wavelengths λ1 to λ6 that are longer than the wavelength λb, while not generating fluorescence having wavelengths λ7 and λ8 that are shorter than the wavelength λb. That is, the light receiving elements 402b-7 to 402b-8 assigned to detect the wavelengths λ7 and λ8 do not need to transmit light. Therefore, a transmission circuit does not need to be connected to the light receiving elements 402b-7 to 402b-8. As a result, signals are not transmitted from the light receiving elements 402b-7 to 402b-8, and the amount of signal transmission can be reduced compared to the case where a transmission circuit is connected to the light receiving elements 402b-7 to 402b-8.

[0065] The photodetector 400c includes a spectroscopic unit 401c, a light receiving element array 402c, and a transmission unit 403c. The photodetector 400c, and the spectroscopic unit 401c and the light receiving element array 402c included therein are the same as the photodetector 300 described in (2-3-1) above, and the spectroscopic unit 301 and the light receiving element array 302, and the description thereof also applies to this example. On the other hand, the transmission unit 403c is different from the transmission unit 303 described in (2-3-1) above, that is, it is also different from the transmission unit 403a of the photodetector 400a. The difference is that a transmission circuit is connected only to some of the light receiving elements included in the light receiving element array 402c, and no transmission circuit is connected to the remaining light receiving elements. More specifically, transmission circuits 403c-1 to 403c-4 are respectively connected to the light receiving elements 402c-1 to 402c-4, while no transmission circuit is connected to the light receiving elements 402c-5 to 402c-8. In this way, the transmission unit 403c is configured not to transmit signals detected by some of the light receiving elements in the light receiving element array 402c.

[0066] The photodetector 400c is associated with a predetermined light source c (not shown) included in the light irradiation unit 2. The light source c emits light (particularly laser light) having a wavelength λc. The wavelength λc is shorter than λ4 and longer than λ5 among the wavelengths λ1 to λ8 of light that can be detected by the light receiving element array 402c. Therefore, the light having the wavelength λc emitted by the light source c may generate fluorescence having wavelengths λ1 to λ4 that are longer than the wavelength λc, while not generating fluorescence having wavelengths λ5 to λ8 that are shorter than the wavelength λc. That is, the light receiving elements 402c-5 to 402c-8 that are assigned to detect wavelengths λ5 to λ8 do not need to transmit light. Therefore, a transmission circuit does not need to be connected to the light receiving elements 402c-5 to 402c-8. As a result, signals are not transmitted from the light receiving elements 402c-5 to 402c-8, and the amount of signal transmission can be reduced as compared with the case where a transmission circuit is connected to the light receiving elements 402c-5 to 402c-8.

[0067] An example in which each of the plurality of photodetectors is controlled so as not to transmit a signal of light having a wavelength equal to or less than the wavelength of the associated light source in this embodiment will be described with reference to FIG. 5. In this example, each photodetector is controlled such that only the light receiving elements that receive light having a wavelength longer than the wavelength of the associated light source among the light receiving element array transmit signals based on the received light.

[0068] The detection unit 3 shown in FIG. 5 includes three photodetectors 500a, 500b, and 500c.

[0069] The photodetector 500a includes a spectroscopic unit 501a, a light receiving element array 502a, and a transmission unit 503a. The photodetector 500a, and the spectroscopic unit 501a, the light receiving element array 502a, and the transmission unit 503a included therein are the same as the photodetector 300, and the spectroscopic unit 301, the light receiving element array 302, and the transmission unit 303 described in (2-3-1) above, and the description thereof also applies to this example. The photodetectors 500b and 500c are also the same as the photodetector 500a.

[0070] Each of the transmission units 503a, 503b, and 503c included in each photodetector is configured such that each of the transmission circuits included therein can be controlled to transmit a signal or not to transmit a signal. For example, the information processing apparatus 100 may control the signal transmission of each transmission circuit.

[0071] For example, assume that a predetermined light source a (not shown) included in the light irradiation unit 2 is associated with the photodetector 500a. The light source a emits light having a wavelength λa (particularly, laser light). The wavelength λa is shorter than any of the wavelengths of the fluorescence λ1 to λ8 detected by the light receiving element array 502a. The light having the wavelength λa emitted by the light source a may generate fluorescence having wavelengths λ1 to λ8 that are longer than the wavelength λa. Therefore, the information processing apparatus 100 can control all of the transmission circuits included in the transmission unit 503a to perform signal transmission.

[0072] A predetermined light source b (not shown) included in the light irradiation unit 2 is associated with the photodetector 500b. The light source b emits light having a wavelength λb (particularly, laser light). The wavelength λb is shorter than λ6 and longer than λ7 among the wavelengths λ1 to λ8 of light detectable by the light receiving element array 502b. Therefore, the light having the wavelength λb emitted by the light source b may generate fluorescence having wavelengths λ1 to λ6 that are longer than the wavelength λb, while not generating fluorescence having wavelengths λ7 and λ8 that are shorter than the wavelength λb. That is, it is not necessary to transmit the light received by the light receiving elements 502b-7 to 502b-8 assigned to detect the wavelengths λ7 and λ8, and the transmission circuits connected to the light receiving elements 502b-7 to 502b-8 do not need to perform signal transmission either. Therefore, for example, by the information processing apparatus 100 controlling the transmission circuits 503b-1 to 503b-6 to perform signal transmission and controlling the transmission circuits 503b-7 to 503b-8 not to perform signal transmission, the signal transmission amount can be reduced as compared with the case where signal transmission is performed for all of the light receiving elements 502b-1 to 502b-8.

[0073] The photodetector 500c is associated with a predetermined light source c (not shown) included in the light irradiation unit 2. The light source c emits light (particularly, laser light) having a wavelength λc. The wavelength λc is shorter than λ4 and longer than λ5 among the wavelengths λ1 to λ8 of light that can be detected by the light receiving element array 502c. Therefore, the light having the wavelength λc emitted by the light source c may generate fluorescence having wavelengths λ1 to λ4 that are longer than λc, while not generating fluorescence having wavelengths λ5 to λ8 that are shorter than λc. That is, there is no need to transmit the light received by the light receiving elements 502c-5 to 502c-8 that are assigned to detect wavelengths λ5 to λ8, and the transmission circuits connected to the light receiving elements 502c-5 to 502c-8 do not need to perform signal transmission. Thus, for example, by controlling the information processing apparatus 100 to control the transmission circuits 503c-1 to 503c-4 to perform signal transmission and control the transmission circuits 503c-5 to 503c-8 not to perform signal transmission, the signal transmission amount can be reduced compared to the case where signal transmission is performed for all of the light receiving elements 502c-1 to 502c-8.

[0074] (2-4) Configuration Example of Optical System

[0075] FIG. 2 shows a schematic configuration example of the optical system in the particle analysis system 1 according to the present technology. As shown in FIG. 2, the particle analysis system 1 includes a light irradiation unit 2 and a detection unit 3.

[0076] The light irradiation unit 2 includes a plurality of laser light sources (LD-1, LD-2, LD-3, ···, and LD-N. Here, N is an arbitrary integer and may be the total number of the first light source and the second light source described above). The plurality of laser light sources include at least one first light source and at least one second light source described in the above (2-1).

[0077] The light irradiation unit 2 includes optical components 601-1, 601-2, 601-3, ··· 601-N that constitute a light guiding optical system. These optical components may be, for example, mirrors or beam splitters and can be appropriately selected according to the configuration of the light guiding optical system. Further, the light irradiation unit 2 may include a lens group (not shown) for condensing and / or homogenizing, for example, laser light. The plurality of laser lights emitted by the plurality of laser light sources are combined by the light guiding optical system and irradiated onto, for example, cells (shown as Sample in FIG. 2).

[0078] The detection unit 3 includes photodetectors (Detection Unit 1, Detection Unit 1, Detection Unit 3, ··· and Detection Unit N. N is an arbitrary integer and may be the total number of the first light source and the second light source described above) associated with each laser light source. Each photodetector may be pre-assigned a laser light source that emits laser light that generates light (particularly fluorescence) to be detected by each photodetector. The detection unit 3 detects the light generated by the light irradiation of the cells by the light irradiation unit 2.

[0079] Based on the light data obtained by the detection unit 3, the information processing device 100 generates spectral data (Spectrum Data-1, Spectrum Data-2, Spectrum Data-3, ··· Spectrum Data-N) as shown on the right side of FIG. 2.

[0080] (2-5) Information Processing Device

[0081] As shown in FIG. 1, the information processing device 100 includes, for example, a processing unit 101 and a storage unit 102.

[0082] The processing unit 101 processes the light data obtained by the light irradiation of the particles by the light irradiation unit. The processing may include unmixing processing. The light data may be, for example, light data including fluorescence data. More specifically, the light data may be light intensity data, and the light intensity may be light intensity data of light including fluorescence.

[0083] The processing unit 101 preferably executes the unmixing process using spectral reference data. By this process, the fluorescence intensity for each fluorescent dye can be obtained from the optical data. Further, by this process, the leakage of fluorescence, which has been a problem in conventional filter-type flow cytometers, is eliminated, and the fluorescence separation performance is improved. In this specification, the spectral reference data (also referred to as SR data) is spectral data of fluorescence generated when predetermined excitation light is irradiated to each phosphor. The SR data can be obtained, for example, by detecting fluorescence generated by irradiating particles labeled with each phosphor alone with predetermined excitation light using a fluorescence detector.

[0084] The spectral reference data used in the unmixing process includes spectral data of fluorescence generated when predetermined excitation light is irradiated to the phosphor labeling the particles. To obtain the spectral reference data used in the unmixing process, for example, first, a particle population to be analyzed is labeled with each of a plurality of phosphors to obtain a plurality of singly stained particle populations. Next, for each of the plurality of singly stained particle populations, spectral data of fluorescence generated by light irradiation (particularly laser light irradiation) is obtained. The obtained spectral data is used as the spectral reference data. In this way, the spectral data of fluorescence regarding the particle populations labeled with each of the plurality of phosphors can be used as the spectral reference data in the unmixing process. Thereby, the fluorescence separation performance can be improved.

[0085] For the light irradiation for obtaining spectral reference data, light (particularly laser light) having the same wavelength as that of the at least one first light source and light (particularly laser light) having the same wavelength as that of the at least one second light source can be used. For example, for the light irradiation for obtaining spectral reference data, the light irradiation unit constituting the particle analysis system of the present technology may be used.

[0086] The unmixing process may be performed according to, for example, the fluorescence intensity correction method or the fluorescence intensity calculation method described in Japanese Patent Application Laid-Open No. 2011-232259 (the above Patent Document 1).

[0087] Examples of the information processing by the processing unit 101 will be described in the following (2-7).

[0088] The storage unit 102 stores various data. The storage unit 102 may be configured to be able to store, for example, the optical data acquired by the detection unit 3. The storage unit 102 may further be configured to be able to store spectral reference data.

[0089] The processing unit 101 can control the output unit 4 to output the processing result of the optical data. Further, the processing unit 101 can receive a signal from the input unit 5 (for example, an operation signal generated by an operation of the input unit 5 by a user), and execute various processes and / or control the information processing apparatus 100 based on the signal.

[0090] A configuration example of the information processing apparatus 100 will be described below. The processing by the processing unit 101 can be realized, for example, by the following configuration, but the configuration of the information processing apparatus 100 is not limited to the following.

[0091] The information processing apparatus 100 may include, for example, a CPU (Central Processing Unit), a RAM, and a ROM. The CPU, the RAM, and the ROM may be connected to each other via a bus. An input / output interface may be further connected to the bus. The output unit 4 and the input unit 5 may be connected to the bus via the input / output interface.

[0092] The input / output interface may further be connected to, for example, a communication device, a storage device, and a drive.

[0093] The communication device connects the information processing device 100 to a network, either wired or wirelessly. By means of the communication device, the information processing device 100 can acquire various data (such as optical data and / or SR data, etc.) via the network. The acquired data can be stored, for example, in the storage unit 102. The type of the communication device may be appropriately selected by those skilled in the art.

[0094] The storage device may store an operating system (such as WINDOWS (registered trademark), UNIX (registered trademark), or LINUX (registered trademark), etc.), a program for causing an information processing device (or a particle analysis device or a particle analysis system) to execute an information processing method according to the present technology, and various other programs, as well as optical data, SR data, and various other data.

[0095] The drive can read data (such as optical data and SR data, etc.) or a program recorded on a recording medium and output it to the RAM. The recording medium is, for example, a microSD memory card, an SD memory card, or a flash memory, but is not limited thereto.

[0096] (2-6) Output unit and input unit

[0097] The output unit 4 includes, for example, a device that outputs the result of the optical data processing by the processing unit 101. For example, the output unit 4 can output fluorescence data obtained by performing an unmixing process or output data generated based on the fluorescence data (for example, a two-dimensional plot based on the fluorescence data). The output unit 4 can include, for example, a display device (display). The display device can output the fluorescence data or output data obtained as a result of the optical data processing as an image (a still image or a moving image). Further, the output unit 4 can include, for example, a printing device. The printing device can print and output the fluorescence data or output data obtained as a result of the optical data processing on a printing medium such as paper.

[0098] The input unit 5 is, for example, a device that receives an operation by a user. The input unit 5 can include, for example, a mouse, a keyboard, or a display (in this case, the user operation can be a touch operation on the display). The input unit 5 transmits an operation by the user to the information processing device 100 as an electrical signal. The processing unit 101 of the information processing device 100 can perform various processes according to the electrical signal.

[0099] (2-7) Example of Information Processing by Information Processing Device

[0100] An example of the information processing by the information processing device 100 will be described below with reference to FIG. 6. FIG. 6 is an example of a flowchart of the information processing by the information processing device 100.

[0101] In step S101, the information processing device 100 starts information processing (particularly, optical data processing). For example, in response to a user clicking a predetermined processing start button displayed on the display of the output unit 4, the processing unit 101 displays a window for performing optical data processing on the display.

[0102] Prior to the start of the processing of the optical data, light detection (particularly flow cytometry) may be performed on the particle population labeled with a plurality of phosphors using the light irradiation unit 2, the chip T, and the detection unit 3 described above, and the optical data obtained as a result may be stored in the storage unit 102.

[0103] In step S102, the processing unit 101 acquires optical data regarding the particle population. The processing unit 101 can receive, for example, the optical data acquired by the detection unit 3. Alternatively, the processing unit 101 may acquire the optical data stored in the storage unit 102.

[0104] In step S103, the processing unit 101 performs an unmixing process on the optical data acquired in step S102. This unmixing process is also called a fluorescence separation process.

[0105] In step S103, the processing unit 101 preferably executes the unmixing process using spectral reference data. The spectral reference data used in the unmixing process includes spectral data of fluorescence generated when a predetermined excitation light is irradiated onto the phosphor labeling the particles. The spectral reference data used in the unmixing process preferably includes spectral data of fluorescence generated when light having the same wavelength as the light emitted by the at least one first light source is irradiated onto the phosphor labeling the particles, and spectral data of fluorescence generated when light having the same wavelength as the light emitted by the at least one second light source is irradiated onto the phosphor labeling the particles.

[0106] The spectral reference data used in step S103 may be stored in the storage unit 102 in advance. The processing unit 101 can acquire the spectral reference data from, for example, the storage unit 102 and perform the unmixing process.

[0107] The processing unit 101 can perform the unmixing process using, for example, the Least Square Method (LSM), more preferably the Weighted Least Square Method (WLSM). The unmixing process using the least square method may be performed using, for example, the fluorescence intensity correction method described in Japanese Patent No. 5985140. The fluorescence intensity correction method can be performed using, for example, the following mathematical formula (1) of WLSM. [Number] In the above mathematical formula (1), x n represents the fluorescence intensity of the n-th fluorescent dye, [S T represents the transposed matrix of the spectral reference, [L] represents the weight matrix, [S] represents the matrix of the spectral reference, y i represents the measured value at the i-th photodetector, λ i represents the weight at the i-th photodetector, max(y i , 0) represents the larger value obtained by comparing the detected value of the i-th detector with zero, and offset’ represents a value determined based on the detected values of each detector.

[0108] The fluorescence wavelength distribution of a phosphor (such as a fluorescent dye) may be broad. Therefore, for example, a PMT used to detect fluorescence generated from a certain phosphor may also detect fluorescence generated from other phosphors. That is, the optical data acquired by each PMT may be data in which fluorescence data from a plurality of phosphors are superimposed. Therefore, correction is required to separate the optical data into fluorescence data from each phosphor. The unmixing process is a technique for such correction, and by the unmixing process, data in which fluorescence data from a plurality of phosphors are superimposed is separated into fluorescence data from each phosphor, and fluorescence data from each phosphor is obtained.

[0109] In step S104, the processing unit 101 generates output data using the fluorescence data obtained by the unmixing process. The output data may be, for example, but is not limited to, a two-dimensional plot regarding two desired phosphors out of a plurality of phosphors used for labeling the particle population. The vertical axis of the two-dimensional plot may be fluorescence data (particularly fluorescence intensity) of fluorescence corresponding to one of the two phosphors, and the horizontal axis may be fluorescence data (particularly fluorescence intensity) of fluorescence corresponding to the other phosphor. The two-dimensional plot may be, for example, a density plot (dot plot), a contour plot, or a plot of both density and contour. The gate setting and unfolding operations for generating the two-dimensional plot may be appropriately performed by the user according to the purpose of particle analysis.

[0110] In step S104, the processing unit 101 can generate one or a plurality (for example, two or more, particularly 2 to 30, more particularly 2 to 20) of generated two-dimensional plots. Also, in step S104, in addition to the two-dimensional plot regarding two phosphors, the processing unit 101 may generate a plot based on scattered light (for example, any two of forward scattered light, side scattered light, and backward scattered light) and / or a plot based on scattered light and fluorescence.

[0111] In step S105, the processing unit 101 can output the output data (for example, a two-dimensional plot, etc.) generated in step S104 to an output unit (for example, a display device of the output unit, etc.).

[0112] In step S106, the processing unit 101 ends the information processing. Also, prior to the end, the processing unit 101 can store the fluorescence data after the unmixing process and / or the generated output data in the storage unit 102.

[0113] (2-8) Particle

[0114] In the present technology, the particles may be particles having dimensions that can flow, for example, within a flow path provided in the chip T. In the present technology, the particles may be appropriately selected by those skilled in the art. In the present technology, the particles may include biological microparticles such as cells, cell aggregates, microorganisms, and liposomes, as well as synthetic microparticles such as gel particles, beads, latex particles, polymer particles, and industrial particles.

[0115] Biological microparticles (also referred to as biological particles) may include chromosomes, liposomes, mitochondria, and organelles (cell organelles) that make up various cells. Cells may include animal cells (such as blood cell line cells) and plant cells. The cells may particularly be blood cell line cells or tissue cell line cells. The blood cell line cells may be, for example, floating cells such as T cells and B cells. The tissue cell line cells may be, for example, adherent cultured cells or adherent cells separated from tissue. Cell aggregates may include, for example, spheroids and organoids. Microorganisms may include bacteria such as Escherichia coli, viruses such as tobacco mosaic virus, and fungi such as yeast. Furthermore, biological microparticles may also include biological macromolecules such as nucleic acids, proteins, and complexes thereof. These biological macromolecules may be, for example, those extracted from cells or may be those contained in a blood sample or other liquid sample. According to one embodiment of the present technology, the particles are biological particles, particularly cells.

[0116] Synthetic microparticles may be, for example, microparticles made of an organic or inorganic polymer material or metal. The organic polymer materials may include polystyrene, styrene divinylbenzene, and polymethyl methacrylate. The inorganic polymer materials may include glass, silica, and magnetic materials. Metals may include gold colloids and aluminum. The synthetic microparticles may be, for example, gel particles or beads, and more particularly may be gel particles or beads to which one or more combinations selected from oligonucleotides, peptides, proteins, and enzymes are bound.

[0117] The shape of the particles may be spherical or substantially spherical, or may be non-spherical. The size and mass of the particles can be appropriately selected by those skilled in the art according to the size of the flow path of the chip. On the other hand, the size of the flow path of the chip can also be appropriately selected according to the size and mass of the particles.

[0118] The particle analysis system of the present technology may be one that analyzes a particle population. In the particle analysis system of the present technology, light irradiation by the light irradiation unit can be performed on each particle included in the particle population. The particle population may particularly be a population of biological particles, and more particularly may be a population of cells.

[0119] The particle analysis system of the present technology may be one that analyzes a particle population labeled with a plurality of phosphors. The phosphor for labeling the particle population may be a fluorescent dye described later. Further, the fluorescent dye may be bound to particles (particularly cells) via a molecule (such as an antibody, aptamer, DNA, or RNA, particularly an antibody) that specifically binds to the particles. The particle population may be contained in the sample solution when light irradiation by the light irradiation unit is performed. The type of the sample solution can be appropriately selected by those skilled in the art and can be determined according to consideration factors such as the type of particles (cells). Note that the type of the sheath fluid can also be appropriately selected by those skilled in the art.

[0120] The plurality of phosphors may be, for example, a plurality of dyes, particularly a plurality of fluorescent dyes. The phosphor may be a phosphor (dye) known in the technical field of flow cytometry, for example. Examples of the fluorescent dye include, but are not limited to, the following: Cascade Blue, Pacific Blue, Fluorescein isothiocyanate (FITC), AleaFluor488, Phycoerythrin (PE), Propidium iodide (PI), Texas Red (TR), PE-efluor 610, PE / Dazzle594, ECD (PE-TxRed), PE-CF594, PE-Vio 615, 7-AAD, PE / Cy5, Peridinin chlorophyll protein (PerCP), PerCP / Cy5.5, PerCP / eFluor710, PE / Cy7, Allophycocyanin (APC), AlexaFluor647, AlexaFluor700, APC-AlexaFluor700, APC / Fire750, APC / eFluor780, APC / H7, Brilliant Violet (BV421), BD Horizon V450, eFluor450, Pacific Blue, 4’,6-Diamidino-2-phenylindole (DAPI), AmCyan, BD Horizon V500, Brilliant Violet 510, PacificOrange, Brilliant Violet 570, Brilliant Violet 605, Brilliant Violet 650, eFluor 650NC, Brilliant Violet 711, Brilliant Violet 785, Cy3, Cy5, and Cy7. Further, the fluorescent dye may be the fluorescent dye described in the examples below.

[0121] The particle population is, for example, a particle population labeled with 5 or more phosphors, more preferably a particle population labeled with 8 or more, 10 or more, 15 or more, or 20 or more phosphors. The particle population may further be a particle population labeled with 22 or more, 24 or more, or 26 or more phosphors. The particle population may be, for example, a particle population labeled with 50 or fewer, 45 or fewer, or 40 or fewer phosphors. The particle analysis system according to the present technology has excellent fluorescence separation performance and is suitable for analyzing a particle population labeled with such a variety of phosphors (fluorescent dyes).

[0122] 2. Second Embodiment (Particle Analyzer)

[0123] The present technology also provides a particle analyzer including a detection unit that detects light generated by irradiating particles flowing in a flow path with light. The detection unit may include at least one photodetector that detects light generated by irradiating the particles with light from the light irradiation unit. Further, the at least one fluorescence detector may have a light receiving element array, and the at least one photodetector may be configured such that a part of the light receiving elements constituting the light receiving element array does not perform signal transmission according to the wavelength of the light, or a part of the light receiving elements constituting the light receiving element array may be controlled not to perform signal transmission. By providing such a detection unit, the particle analyzer of the present technology can reduce the amount of signal transmission.

[0124] The detection unit is as described in (2-3) above, and the description thereof also applies to the present embodiment.

[0125] In addition to the detection unit, the particle analyzer of the present technology may include the light irradiation unit described in (2-1) above.

[0126] The particle analyzer of the present technology may be configured as a particle analysis system in combination with the information processing device described in (2-5) above and the output unit and input unit described in (2-6) above. Alternatively, the particle analyzer of the present technology may include any one or more of the information processing device, the output unit, and the input unit.

[0127] 3. Third Embodiment (Information Processing Method)

[0128] The present technology also provides an information processing method including an unmixing process of performing an unmixing process on light data obtained by irradiating particles with light from a light irradiation unit including at least one first light source that emits light having a wavelength of 350 nm or more and at least one second light source that emits light having a wavelength of less than 350 nm.

[0129] The information processing method according to the present technology can be performed, for example, to process data obtained from the results of fluorescence detection of a particle population labeled with a plurality of phosphors. The information processing method according to the present technology may be executed, for example, by the particle analysis system described in 1. above or the particle analyzer described in 2. above, and particularly may be executed by an information processing device that may be included in the particle analysis system or the particle analyzer.

[0130] The information processing method according to the present technology may include, for example, a light data acquisition step of acquiring light data about a particle population, an unmixing process step of performing an unmixing process on the acquired light data, an output data generation step of generating output data using the fluorescence data after the unmixing process, and a data output step of outputting the generated output data.

[0131] The light data acquisition step corresponds to step S102 described in (2-7) of 1. above. The unmixing process step corresponds to step S103 described in (2-7) of 1. above. The output data generation step corresponds to step S104 described in (2-7) of 1. above. The data output step corresponds to step S105 described in (2-7) of 1. above. Therefore, the descriptions of these steps in (2-7) of 1. above also apply to each step in the information processing method of the present technology.

[0132] 4. Fourth Embodiment (Program)

[0133] The present technology also provides a program for causing an information processing apparatus to execute the information processing method described in 3. above. The information processing method is as described in 1. and 3. above, and this description also applies to the present embodiment. The program according to the present technology may be recorded, for example, on the recording medium described above, or may be stored in the information processing apparatus or the storage device included in the information processing apparatus described above.

[0134] 5. Examples

[0135] Particle analysis processing was performed using a flow cytometer equipped with a light irradiation unit including a laser light source that emits laser light with a wavelength of 320 nm and six laser light sources that emit laser light with a wavelength of 350 nm or more.

[0136] As an analysis target for the particle analysis processing, a cell population obtained by hemolyzing human normal blood was used as a sample.

[0137] In addition, 43 antibodies each conjugated with the following 43 fluorescent dyes were prepared. All of the prepared antibodies were anti-CD4 antibodies. Alexa Fluor 488, Alexa Fluor 514, Alexa Fluor 647, Alexa Fluor 700, APC-Cy7, APC, BUV395, BUV496, BUV563, BUV661, BUV737, BUV805, BV421, BV480, BV510, BV570, BV605, BV650, BV711, BV750, BV786, Cy3, PacificBlue, PacificOrange, PE-AF610, PE-AF700, PE-Cy5, PE-Cy5.5, PE-Cy7, PE-Dazzle594, PE, PerCP-Cy5.5, PerCP-eF710, PerCP, Qdot 525, Qdot 545, Qdot 565, Qdot 585, Qdot 605, Qdot 625, Qdot 655, Qdot 705, and SB702

[0138] Each of the 43 fluorescent dye-conjugated antibodies was used to label the sample. As a result, 43 samples labeled with each antibody were obtained. The 43 obtained samples were each placed in a tube. Also, an unlabeled sample that was not labeled with any antibody was prepared in a tube. As a result, a total of 44 sample-containing tubes were obtained.

[0139] For each of the 44 samples, 44 fluorescence spectrum data were acquired using the flow cytometer. The 44 fluorescence spectrum data were used as spectral reference data below.

[0140] Also, the 44 fluorescence spectrum data were merged to obtain merged data. The merged data were subjected to unmixing processing using the 44 spectral reference data. Using the data obtained by the unmixing processing, a two-dimensional plot was generated. The generated two-dimensional plot is shown in FIG. 7.

[0141] Except for not turning on the laser light source that emits laser light with a wavelength of 320 nm, for each of the 44 samples, 44 pieces of fluorescence spectrum data were acquired using the flow cytometer in the same manner as above. The 44 pieces of fluorescence spectrum data were used as spectral reference data below. Also, the 44 pieces of fluorescence spectrum data were merged in the same manner as above to obtain merged data. The merged data was subjected to unmixing processing in the same manner as above using the 44 pieces of spectral reference data, and a two-dimensional plot was generated. The generated two-dimensional plot is shown in FIG. 8.

[0142] From the comparison of the two-dimensional plots of FIGS. 7 and 8, it can be seen that the fluorescence separation performance was improved by using laser light with a wavelength of 320 nm. Also, the S / N ratio when using laser light with a wavelength of 320 nm was improved by about 30% compared to the case where the laser light with a wavelength of 320 nm was not lit.

[0143] From the above results, it can be seen that in the fluorescence analysis of particles, by using a laser light source that emits laser light with a wavelength of less than 350 nm in combination with six laser light sources that emit laser light with a wavelength of 350 nm or more, and performing unmixing processing, the fluorescence separation performance is improved.

[0144] Note that the present technology can also have the following configuration. 〔1〕 An optical irradiation unit including at least one first light source that emits light with a wavelength of 350 nm or more and at least one second light source that emits light with a wavelength of less than 350 nm, and A processing unit that performs unmixing processing on the optical data obtained by irradiating the particles with the light from the optical irradiation unit. A particle analysis system including the above. 〔2〕 The particle analysis system according to 〔1〕, wherein the at least one second light source emits light with a wavelength of 250 nm or more and less than 350 nm. 〔3〕 The particle analysis system according to [1] or [2], wherein at least two of the excitation lights emitted by the at least one first light source and the excitation lights emitted by the at least one second light source are combined, and the combined excitation light is configured to irradiate particles. 〔4〕 The particle analysis system according to any one of [1] to [3], which analyzes a particle population labeled with a plurality of phosphors. 〔5〕 The particle analysis system according to any one of [1] to [4], wherein the processing unit performs the unmixing process using spectral reference data. 〔6〕 The particle analysis system according to [4], wherein spectral data of fluorescence related to a particle population labeled with each of the plurality of phosphors is used as the spectral reference data used in the unmixing process. 〔7〕 The particle analysis system according to any one of [1] to [6], further including a detection unit that detects light generated by irradiating particles with light from the light irradiation unit. 〔8〕 The detection unit includes at least one photodetector that detects light generated by irradiating particles with light from the light irradiation unit, The at least one photodetector has a light receiving element array, and The processing unit acquires only a signal based on light received by a part of the light receiving elements constituting the light receiving element array according to the wavelength of the light. The particle analysis system according to [7]. 〔9〕 The at least one photodetector is configured such that a part of the light receiving elements constituting the light receiving element array does not perform signal transmission according to the wavelength of the light. The particle analysis system according to [8]. 〔10〕 The at least one photodetector is controlled such that a part of the light receiving elements constituting the light receiving element array does not perform signal transmission according to the wavelength of the light. The particle analysis system according to [8].

[11] The detection unit includes a plurality of photodetectors, Each of the plurality of photodetectors is associated with a light source included in the light irradiation unit, The processing unit acquires only a signal based on light received by a part of the light receiving elements constituting the light receiving element array of each photodetector according to the wavelength of the light source associated with each photodetector. The particle analysis system according to [7].

[12] Each of the plurality of photodetectors is configured not to transmit a signal of light having a wavelength equal to or less than the wavelength of the associated light source, and Among the light receiving elements constituting the light receiving element array of each photodetector, only the light receiving elements that receive light having a wavelength longer than the wavelength of the associated light source are connected to a signal transmission circuit that transmits a signal based on the received light. The particle analysis system according to

[11] .

[13] Each of the plurality of photodetectors is controlled not to transmit a signal of light having a wavelength equal to or less than the wavelength of the associated light source, and Each photodetector is controlled such that only the light receiving elements that receive light having a wavelength longer than the wavelength of the associated light source among the light receiving elements constituting the light receiving element array transmit a signal based on the received light. The particle analysis system according to

[11] .

[14] The plurality of photodetectors have the same light receiving element array. The particle analysis system according to any one of

[11] to

[13] .

[15] The particle analysis system according to any one of [1] to

[14] , wherein the at least one first light source is a laser light source and the at least one second light source is a laser light source.

[16] An unmixing processing step of unmixing optical data obtained by irradiating particles with light from an optical irradiation unit including at least one first light source that emits light with a wavelength of 350 nm or more and at least one second light source that emits light with a wavelength of less than 350 nm. Information processing method. 〔17〕 A program for causing an information processing apparatus to execute an unmixing processing step of unmixing optical data obtained by irradiating particles with light from an optical irradiation unit including at least one first light source that emits light with a wavelength of 350 nm or more and at least one second light source that emits light with a wavelength of less than 350 nm.

Explanation of symbols

[0145] 1 Particle analysis system 2 Optical irradiation unit T Chip 3 Detection unit 100 Information processing apparatus 101 Processing unit 102 Storage unit 4 Output unit 5 Input unit

Claims

1. An optical irradiation unit including at least one first light source that emits light having a wavelength of 350 nm or more and at least one second light source that emits light having a wavelength of less than 350 nm, and A processing unit that performs an unmixing process on the optical data obtained by irradiating the particles with light from the optical irradiation unit, A detection unit that detects the light generated by irradiating the particles with light from the optical irradiation unit, comprising, The detection unit includes a plurality of photodetectors, Each of the plurality of photodetectors is associated with a light source included in the optical irradiation unit, The processing unit acquires only a signal based on light received by a part of the light receiving elements constituting the light receiving element array of each photodetector according to the wavelength of the light source associated with each photodetector. A particle analysis system.

2. The particle analysis system according to claim 1, wherein the at least one second light source emits light having a wavelength of 250 nm or more and less than 350 nm.

3. The particle analysis system according to claim 1, wherein at least two of the excitation lights emitted by the at least one first light source and the excitation lights emitted by the at least one second light source are combined, and the combined excitation light is configured to irradiate the particles.

4. The particle analysis system according to claim 1, wherein the particle analysis system analyzes a particle population labeled with a plurality of phosphors.

5. The particle analysis system according to claim 1, wherein the processing unit performs the unmixing process using spectral reference data.

6. The particle analysis system according to claim 4, wherein the spectral data of the fluorescence related to the particle population labeled with each of the plurality of phosphors is used as the spectral reference data used in the unmixing process.

7. The plurality of photodetectors are controlled such that, according to the wavelength of the light, some of the light-receiving elements constituting the light-receiving element array do not perform signal transmission. The particle analysis system according to claim 1.

8. Each of the plurality of photodetectors is configured not to transmit a signal of light having a wavelength equal to or less than the wavelength of the associated light source, and Among the light-receiving elements constituting the light-receiving element array of each photodetector, only the light-receiving elements that receive light having a wavelength longer than the wavelength of the associated light source are connected to a signal transmission circuit that transmits a signal based on the received light. The particle analysis system according to claim 1.

9. Each of the plurality of photodetectors is controlled not to transmit a signal of light having a wavelength equal to or less than the wavelength of the associated light source, and Each photodetector is controlled such that, among the light-receiving elements constituting the light-receiving element array, only the light-receiving elements that receive light having a wavelength longer than the wavelength of the associated light source transmit a signal based on the received light. The particle analysis system according to claim 1.

10. The plurality of photodetectors have the same light-receiving element array. The particle analysis system according to claim 1.

11. The particle analysis system according to claim 1, wherein the at least one first light source is a laser light source and the at least one second light source is a laser light source.

12. Irradiate particles with light from a light irradiation unit including at least one first light source that emits light having a wavelength of 350 nm or more and at least one second light source that emits light having a wavelength of less than 350 nm, including an unmixing process step of detecting the light generated by irradiating the particles with the light irradiation and performing an unmixing process on the obtained light data by a plurality of photodetectors. Each of the plurality of photodetectors is associated with a light source included in the light irradiation unit. The unmixing process is performed by acquiring only a signal based on light received by a part of the light receiving elements constituting the light receiving element array of each photodetector according to the wavelength of the light source associated with each photodetector. Information processing method.

13. Irradiate particles with light using a light irradiation unit including at least one first light source that emits light with a wavelength of 350 nm or more and at least one second light source that emits light with a wavelength of less than 350 nm. Detect the light generated by irradiating the particles with the light irradiation by a plurality of photodetectors, and perform an unmixing process on the obtained light data. Each of the plurality of photodetectors is associated with a light source included in the light irradiation unit. A program for causing an information processing apparatus to execute an unmixing process step, wherein the unmixing process is performed by acquiring only a signal based on light received by a part of the light receiving elements constituting the light receiving element array of each photodetector according to the wavelength of the light source associated with each photodetector.

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