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US-20260126368-A1 - SPECTRUM ANALYSIS APPARATUS, FINE PARTICLE MEASUREMENT APPARATUS, AND METHOD AND PROGRAM FOR SPECTRUM ANALYSIS OR SPECTRUM CHART DISPLAY

US20260126368A1US 20260126368 A1US20260126368 A1US 20260126368A1US-20260126368-A1

Abstract

Provided is a spectrum analysis apparatus including a processing unit configured to generate analysis data using an analysis function in which a linear function and a logarithmic function are included as function elements and an intensity value is set as a variable from measurement data including the intensity value of light acquired by detecting the light from a measurement target object using a plurality of light-receiving elements having different detection wavelength bands.

Inventors

  • Nao Nitta

Assignees

  • SONY CORPORATION

Dates

Publication Date
20260507
Application Date
20251229
Priority Date
20110913

Claims (20)

  1. 1 . A spectral flow cytometer system comprising: a light source configured to irradiate light with a first wavelength region to a plurality of particles stained with at least one of a plurality of fluorescent dyes; a spectral element configured to separate light emitted from the plurality of particles; an optical filter that filters light separated by the spectral element to limit the light within the first wavelength region; a multi-channel detector configured to detect light filtered by the optical filter, each channel respectively corresponding to different wavelength regions; and a processor configured to receive a signal from the multi-channel detector and generate analysis data.
  2. 2 . The spectral flow cytometer system of claim 1 , wherein the spectral element comprises a grating or prism.
  3. 3 . The spectral flow cytometer system of claim 2 , wherein the spectral element comprises a grating.
  4. 4 . The spectral flow cytometer system of claim 2 , wherein the spectral element comprises a prism.
  5. 5 . The spectral flow cytometer system of claim 1 , wherein the light source comprises a plurality of light sources and the spectral element comprises a plurality of spectral elements.
  6. 6 . The spectral flow cytometer system of claim 1 , wherein the light source is further configured to irradiate the excitation wavelength through a flow path configured to flow the particles through the flow cytometer system.
  7. 7 . The spectral flow cytometer system of claim 1 , wherein the plurality of particles comprise unlabeled particles and labeled particles.
  8. 8 . The spectral flow cytometer system of claim 1 , wherein the plurality of fluorescent dyes is less than a number of the channels in the multi-channel detector in the spectral flow cytometer apparatus.
  9. 9 . The spectral flow cytometer system of claim 1 , wherein the multi-channel detector is further configured to detect light over a range of 500 nm to 800 nm.
  10. 10 . The spectral flow cytometer system of claim 1 , wherein the processor is further configured to generate the analysis data using: (i) a linear function to assess whether the signal from the multi-channel detector has an intensity value below a set value; and (ii) a logarithmic function of an analysis function to assess whether the signal from the multi-channel detector has an intensity value above the set value.
  11. 11 . The spectral flow cytometer system of claim 1 , further comprising a display unit configured to display the analysis data in a spectrum chart.
  12. 12 . The spectral flow cytometer system of claim 11 , wherein the spectrum chart comprises a first axis and a second axis, where the first axis represents a value corresponding to the plurality of wavelength regions and the second axis represents an intensity value.
  13. 13 . The spectral flow cytometer system of claim 11 , wherein the spectrum chart comprises an axis of an intensity value set as a logarithmic axis in a region in which the intensity value is greater than a predetermined value, and wherein the spectrum chart comprises an axis of an intensity value set as a linear axis in a region in which the intensity value is less than a determined value.
  14. 14 . The spectral flow cytometer system of claim 11 , wherein the spectrum chart includes at least one negative value within a linear axis of an intensity value.
  15. 15 . A method for detecting and displaying a spectrum chart comprising: irradiating, by a light source in a spectral flow cytometer apparatus, light with a first wavelength region to a plurality of particles, wherein the particles include cells flowing through a flow path in the spectral flow cytometer apparatus stained with at least one of a plurality of fluorescent dyes, separating, by a spectral element, in the spectral flow cytometer apparatus, light emitted from the plurality of particles; filtering, by an optical filter in the spectral flow cytometer apparatus, light separated by the spectral element to limit the light within the first wavelength region; detecting, by a multi-channel detector in the spectral flow cytometer apparatus, light filtered by the optical filter, each channel respectively corresponding to different wavelength regions; receiving and processing, by a processor in the spectral flow cytometer apparatus, a signal from the multi-channel detector and generating analysis data; and displaying, by a display in the spectral flow cytometer apparatus, a spectrum chart representing analysis data corresponding to each intensity value of the detected light the different wavelength regions, wherein the plurality of fluorescent dyes is less than a number of the channels in the multi-channel detector in the spectral flow cytometer apparatus.
  16. 16 . The method according to claim 15 , further comprising, preventing, by the optical filter, leakage of light from the light source.
  17. 17 . The method according to claim 15 , in the spectrum chart, one axis represents a value corresponding to the plurality of wavelength regions and the other axis represents the intensity value.
  18. 18 . The method according to claim 15 , wherein the multi-channel detector comprises a light-receiving element array of a plurality of detection channels having different detection wavelength bands.
  19. 19 . The method according to claim 15 , wherein the plurality of particles comprise unlabeled particles and labeled particles.
  20. 20 . The method according to claim 19 , further comprising: correcting, by a processor, a measurement value detected by the labeled particles using a measurement value detected by unlabeled particles.

Description

CROSS REFERENCES TO RELATED APPLICATIONS This application is a continuation of U.S. patent application Ser. No. 19/028,862, filed Jan. 17, 2025, which is a continuation of U.S. patent application Ser. No. 16/996,263, filed on Aug. 18, 2020, now U.S. Pat. No. 12,276,591, which is a continuation of U.S. patent application Ser. No. 14/342,587, filed on Mar. 4, 2014, now U.S. Pat. No. 10,782,224, which is a national stage application of International Application No. PCT/JP2012/005780, filed on Sep. 12, 2012 and claims priority to Japanese Patent Application No. 2011-199901, filed on Sep. 13, 2011, each of which is incorporated herein by reference in its entirety. BACKGROUND The present technology relates to a spectrum analysis apparatus, a fine particle measurement apparatus, and a method and program for a spectrum analysis or a spectrum chart display. More particularly, the present technology relates to a spectrum analysis apparatus and the like capable of obtaining a spectrum chart accurately reflecting optical characteristics of a measurement target object. A flow cytometer is an apparatus that optically measures characteristics of fine particles by radiating light to the fine particles such as cells, beads, or the like that flow through a flow cell and detecting fluorescence, scattered light, or the like emitted from the fine particles. For example, when the fluorescence of cells is detected, excitation light having an appropriate wavelength and intensity such as laser light is radiated to a cell labeled by a fluorochrome. The fluorescence emitted from the fluorochrome is condensed by a lens or the like, light of an appropriate wavelength band is selected using a wavelength selection element such as a filter or a dichroic mirror, and the selected light is detected using a light-receiving element such as a photo multiplier tube (PMT). At this time, it is possible to simultaneously detect and analyze fluorescence from a plurality of fluorochromes labeled to cells by a plurality of combinations of wavelength selection elements and light-receiving elements. Further, it is also possible to increase the number of analyzable fluorochromes by combining excitation light of a plurality of wavelengths. In the related art, analysis data of the flow cytometer is displayed by a histogram or a two-dimensional (2D) plot. Although a linear axis or a logarithmic axis is generally used as a coordinate axis representing an intensity value of light in the histogram and the 2D plot, technology using a biexponential axis having characteristics in which the linear axis and the logarithmic axis are combined is also known (see NPL 1). In the histogram and the 2D plot using the biexponential axis as the coordinate axis, a display of a wide dynamic range utilizing characteristics of the logarithmic axis is possible and simultaneously a display of a negative number according to characteristics of the linear axis is also possible. In the fluorescence detection by the flow cytometer, there is a method of measuring an intensity of light in a continuous wavelength band as a fluorescence spectrum in addition to a method of selecting a plurality of pieces of light of a discontinuous wavelength band using a wavelength selection element such as a filter and measuring an intensity of light of each wavelength band. In a spectral flow cytometer in which a fluorescence spectrum is measurable, the fluorescence emitted from the fine particles is spectrally separated using a spectral element such as a prism or a grating. The spectrally separated fluorescence is detected using a light-receiving element array in which a plurality of light-receiving elements having different detection wavelength bands are arranged. As the light-receiving element array, a PMT array or a photodiode array in which light-receiving elements such as PMTs or photodiodes are arranged in one dimension or an array of a plurality of independent detection channels of 2D light-receiving elements such as charge-coupled devices (CCDs) or complementary metal-oxide-semiconductors (CMOSs) is used. CITATION LIST Patent Literature PTL 1: JP 2003-83894A Non Patent Literature NPL 1: A New “Logicle” Display Method Avoids Deceptive Effects of Logarithmic Scaling for Low Signals and Compensated Data. Cytometry Part A 69A: 541-551, 2006. SUMMARY Technical Problem The analysis data in the spectral flow cytometer can be displayed by a spectrum chart in addition to the histogram and the 2D plot. In the spectrum chart, a channel or a detection wavelength of the light-receiving element is represented on the horizontal axis, an intensity value of light is represented on the vertical axis, and information (population information) regarding the number of fine particles (an event count or density) is represented by the gradation of color, a color tone, or the like. According to the spectrum chart, it is possible to intuitively recognize a fluorescence spectrum and population information of fine particles