Digitally enhanced microscopy for multiplexed histology

Patent metadata

Field	Value
Publication number	US-11070750-B2
Application number	US-202016941351-A
Country	US
Kind code	B2
Filing date	Jul 28, 2020
Priority date	Mar 12, 2013
Publication date	Jul 20, 2021
Grant date	Jul 20, 2021

Abstract

Disclosed herein are embodiments of imaging biological specimens. An imaging system can include a microscope for directly viewing the biological specimen and a multi-spectral imaging apparatus for outputting digitally enhanced images, near-video rate imaging, and/or videos of the specimen. An imaging system can include a digital scanner that digitally processes images to produce a composite image with enhanced color contrast of features of interest.

First claim

What is claimed is:

1. A system for generating a multicolor video of a sample on a microscope slide, comprising:
at least one processor; and
memory storing program instructions that, when executed by the at least one processor, cause the system to:
cause a plurality of light sources to output different light wavebands at different times to sequentially illuminate the sample, wherein the plurality of light sources includes one or more of an ultraviolet light source or an infrared light source,
receive a first image of the sample exposed to a first light waveband from a first light source of the plurality of light sources, wherein the first light waveband is configured to interact with a first chromogen bound to a first component of the sample,
receive a second image of the sample exposed to a second light waveband from a second light source of the plurality of light sources, wherein the second light waveband is configured to interact with a second chromogen bound to a second component of the sample, the second light waveband being different from the first light waveband,
generate an enhanced color composite image of the sample based on the first and second images by selecting a false color for at least one of the first or second chromogens, such that color contrast between the first and second components in the enhanced color composite image is increased compared to natural color contrast between the first and second components in the first and second images, and
output the enhanced color composite image as part of a real-time video of the sample.

2. The computer-based imaging system of claim 1 , wherein the plurality of light sources includes both the ultraviolet light source and the infrared light source.

3. The system of claim 1 , wherein the first light source is the ultraviolet light source or the infrared light source.

4. The system of claim 1 , further comprising:
an illuminator including the plurality of light sources; and
an imaging device configured to capture the first and second images.

5. The system of claim 4 , wherein the plurality of light sources are pulsed light sources, and wherein the imaging device is synchronized with pulsing of the pulsed light sources.

6. The system of claim 4 , wherein the plurality of light sources include a plurality of LEDs.

7. The system of claim 1 , wherein:
in the first image, the sample is exposed only to the first light waveband from the first light source; and
in the second image, the sample is exposed only to the second light waveband from the second light source.

8. The system of claim 1 , wherein the instructions further cause the system to convert one or more of the first image or the second image into a false color image.

9. The system of claim 8 , wherein the instructions further cause the system to:
convert the first image to a first false color image; and
convert the second image to a second false color image.

10. The system of claim 9 , wherein the enhanced color composite image is generated by combining the first and second false color images.

11. The system of claim 1 , wherein the instructions further cause the system to capture at least two additional images of the sample exposed to at least two additional light wavebands, wherein the enhanced color composite image is generated based on the first image, the second image, and the at least two additional images.

12. The system of claim 11 , wherein the first image, the second image, and the at least two additional images are each converted into false color images.

13. A method for producing a multicolor video of a sample, the method comprising:
capturing a first image of the sample exposed to a first light waveband from a first light source, the first light waveband being an ultraviolet waveband or an infrared waveband, wherein the sample includes a first feature stained with a first chromogen configured to absorb the first light waveband;
capturing a second image of the sample exposed to a second light waveband from a second light source, the second light waveband being different from the first light waveband, wherein the sample includes a second feature stained with a second chromogen configured to absorb the second light waveband;
producing a digitally-enhanced multicolor image of the sample from the first and second images by converting at least one of the first or second images into a false color image, such that color contrast between the first and second features in the digitally-enhanced multicolor image is increased compared to natural color contrast between the first and second features in the first and second images, and
outputting a multicolor video including the digitally-enhanced multicolor image, wherein the multicolor video is configured to be displayed at a frame rate greater than or equal to 2 frames per second.

14. The method of claim 13 , wherein the first image is captured while the sample is not exposed to the second light waveband from the second light source, and the second image is captured while the sample is not exposed to the first light waveband from the first light source.

15. The method of claim 13 , wherein the capturing of the first and second images is performed without using filters.

16. The method of claim 13 , wherein the first and second light sources are different pulsed light sources, and wherein capturing the first and second images includes sequentially pulsing the different pulsed light sources.

17. The method of claim 16 , wherein the different pulsed light sources are different color LEDs.

18. The method of claim 13 , wherein the multicolor video is configured to be displayed at a frame rate greater than or equal to 30 frames per second.

19. The method of claim 13 , wherein the first chromogen is covalently bound to the first feature and the second chromogen is covalently bound to the second feature.

20. The method of claim 13 , further comprising capturing two or more additional images of the sample exposed to two or more additional light wavebands, wherein the digitally-enhanced multicolor image is produced based on the first image, the second image, and the two or more additional images.

Assignees

• Ventana Med Syst Inc

Inventors

• Morrison Larry
• Dietz Lou

Classifications

• H04N23/11 Primary

  for generating image signals from visible and infrared light wavelengths · CPC title

• G01N21/6458 Primary

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

• H04N5/332 Primary

  Electricity · mapped topic