Fluorescence observation apparatus and fluorescence observation method

Patent metadata

Field	Value
Publication number	US-11662316-B2
Application number	US-201917058091-A
Country	US
Kind code	B2
Filing date	May 30, 2019
Priority date	May 30, 2018
Publication date	May 30, 2023
Grant date	May 30, 2023

Abstract

A fluorescence observation apparatus according to an embodiment of the present technology includes a stage, an excitation section, and a spectroscopic imaging section. The stage is capable of supporting a fluorescently stained pathological specimen. The excitation section irradiates the pathological specimen on the stage with a plurality of line illuminations of different wavelengths, the plurality of line illuminations being a plurality of line illuminations situated on different axes and parallel to a certain-axis direction. The spectroscopic imaging section includes at least one imaging device capable of separately receiving pieces of fluorescence respectively excited with the plurality of line illuminations.

First claim

The invention claimed is:

1. A fluorescence observation apparatus, comprising:
a stage that is capable of supporting a fluorescently stained pathological specimen;
an excitation section that irradiates the pathological specimen on the stage with a plurality of line illuminations of different wavelengths, the plurality of line illuminations being a plurality of line illuminations situated on different axes and being collimated by a collimator to a certain-axis direction; and
a spectroscopic imaging section that includes at least one imaging device capable of separately receiving pieces of fluorescence respectively excited with the plurality of line illuminations, wherein optical axes of the plurality of line illuminations are parallel between an objective lens and the pathological specimen and wherein optical axes of the separately-received pieces of fluorescence are parallel when incident on the spectroscopic imaging section.

2. The fluorescence observation apparatus according to claim 1 , wherein
the excitation section is configured to irradiate, onto the pathological specimen and as the plurality of line illuminations, a plurality of line illuminations each having different wavelengths in combination.

3. The fluorescence observation apparatus according to claim 2 , wherein
the spectroscopic imaging section further includes a wavelength dispersion element that separates each of the pieces of fluorescence respectively excited with the plurality of line illuminations.

4. The fluorescence observation apparatus according to claim 1 , wherein
the spectroscopic imaging section further includes an observation slit including a plurality of slit portions, each of the pieces of fluorescence respectively excited with the plurality of line illuminations being allowed to pass through a corresponding one of the plurality of slit portions.

5. The fluorescence observation apparatus according to claim 1 , further comprising
a scanning mechanism that scans the plurality of line illuminations over the stage in a direction orthogonal to the certain-axis direction.

6. The fluorescence observation apparatus according to claim 1 , further comprising
a processing unit that includes a storage that stores therein spectroscopic data that indicates a correlation between a wavelength of each of the plurality of line illuminations and fluorescence received by the imaging device.

7. The fluorescence observation apparatus according to claim 6 , wherein
the processing unit further includes an image formation section that forms a fluorescence image of the pathological specimen on a basis of the spectroscopic data stored in the storage and an interval between the plurality of line illuminations.

8. The fluorescence observation apparatus according to claim 7 , wherein
the image formation section forms, as the fluorescence image, an image in which coordinates detected by the imaging device have been corrected using a value corresponding to the interval between the plurality of line illuminations.

9. The fluorescence observation apparatus according to claim 6 , wherein
the processing unit further includes a data calibration section that calibrates spectroscopic data stored in the storage.

10. The fluorescence observation apparatus according to claim 7 , wherein
the storage stores therein standard spectra in advance, the standard spectra being a standard spectrum of autofluorescence related to the pathological specimen and a standard spectrum of only a dye that stains the pathological specimen, and
the image formation section outputs a component distribution of the spectroscopic data on a basis of the standard spectrum of the autofluorescence and the standard spectrum of only the dye.

11. The fluorescence observation apparatus according to claim 4 , wherein
the imaging device includes a plurality of imaging devices each capable of receiving the fluorescence passing through the observation slit.

12. The fluorescence observation apparatus according to claim 1 , further comprising:
a non-fluorescence observation section that includes a light source that illuminates the pathological specimen on the stage; and
an imaging section that acquires a non-fluorescence image of the pathological specimen.

13. The fluorescence observation apparatus according to claim 1 , further comprising:
a display section that displays fluorescence spectra separately for each of the plurality of line illuminations, the fluorescence spectra being respectively excited with the plurality of line illuminations.

14. The fluorescence observation apparatus according to claim 13 , wherein
the display section includes an operation region in which a wavelength and output of each of the plurality of line illuminations are allowed to be set.

15. The fluorescence observation apparatus according to claim 13 , wherein
the display section includes a display region used to display a detection wavelength range for the fluorescence spectrum.

16. A fluorescence observation method, comprising:
irradiating a pathological specimen on a stage with a plurality of line illuminations of different wavelengths, the plurality of line illuminations being a plurality of line illuminations situated on different axes and being collimated by a collimator to a certain-axis direction; and
separately receiving pieces of fluorescence respectively excited with the plurality of line illuminations, wherein optical axes of the plurality of line illuminations are parallel between an objective lens and the pathological specimen and wherein optical axes of the separately-received pieces of fluorescence are parallel when incident on a spectroscopic imaging section.

17. The fluorescence observation method according to claim 16 , further comprising scanning the plurality of line illuminations over the stage in a direction orthogonal to the certain-axis direction.

18. The fluorescence observation method according to claim 16 , wherein
a plurality of line illuminations each having different wavelengths in combination is used as the plurality of line illuminations.

Assignees

• Sony Corp

Inventors

• Tatsuta Hirokazu
• Hashimoto Gakuji
• Kuwayama Tetsuro

Classifications

• G01N21/6458 Primary

  Fluorescence microscopy (fluorescence microscopes per se G02B21/0076 and G02B21/16) · CPC title

• G02B21/0064

  multi-spectral or wavelength-selective arrangements, e.g. wavelength fan-out, chromatic profiling (G02B21/0076 takes precedence) · CPC title

• G02B21/367

  providing an output produced by processing a plurality of individual source images, e.g. image tiling, montage, composite images, depth sectioning, image comparison · CPC title

• G02B27/1006

  for splitting or combining different wavelengths (G02B27/1086, G02B27/141 take precedence) · CPC title

• G01N2201/12761

  Precalibration, e.g. for a given series of reagents · CPC title