# US 2014/0193061 A1 · Whole Slide Multispectral Imaging Systems and Methods > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 2014/0193061 A1 - Document ID: US20140193061A1 - Status: Published application - Kind: A1 - Application number: 14/149,020 - Filed: 2014-01-07 - Published: 2014-07-10 - Earliest claimed: 2013-01-10 - Assignees: Akoya Biosciences, Inc. - Applicants: Caliper Life Sciences, Inc. - Inventors: Peter J. Miller - Web page: https://patentgrep.com/patent/US20140193061A1 ## CPC - G06T 7/0012: {Biomedical image inspection} - G06T 7/32: using correlation-based methods - G06T 7/97: {Determining parameters from multiple pictures (depth or shape recovery from multiple images G06T7/55; stereo camera calibration G06T7/85)} - G06T 2207/10036: Multispectral image; Hyperspectral image - G06T 2207/10056: Microscopic image - G06T 2207/10064: Fluorescence image - G06T 2207/20212: Image combination - G06T: IMAGE DATA PROCESSING OR GENERATION, IN GENERAL ## Abstract The methods and systems disclosed herein include obtaining a first plurality of images of a sample, where each image in the first plurality of images corresponds to a different spectral band of illumination light incident on the sample or emission light from the sample, obtaining a second plurality of images of the sample, where each image in the second plurality of images corresponds to a different spectral band of illumination light incident on the sample or emission light from the sample, aligning the first and second pluralities of images based on information from a first image from the first plurality of images and a second image from the second plurality of images, where the first and second images correspond to a shared spectral band, and combining at least some members of the first plurality of images and at least some members of the second plurality of images to form an image stack. ## Claims ### Claim 1 A method, comprising: obtaining a first plurality of images of a sample, wherein each image in the first plurality of images corresponds to a different spectral band of illumination light incident on the sample or emission light from the sample; obtaining a second plurality of images of the sample, wherein each image in the second plurality of images corresponds to a different spectral band of illumination light incident on the sample or emission light from the sample; and using an electronic processor to: align the first and second pluralities of images based on information from a first image from the first plurality of images and a second image from the second plurality of images, wherein the first and second images correspond to a shared spectral band; and combine at least some members of the first plurality of images and at least some members of the second plurality of images to form an image stack. ### Claim 2 (depends on 1) The method of claim 1, wherein each pixel in the image stack comprises spectral information derived from at least one member of the first plurality of images and spectral information derived from at least one member of the second plurality of images. ### Claim 3 (depends on 1) The method of claim 1, wherein aligning the first and second pluralities of images comprises using the electronic processor to spatially register the first and second pluralities of images so that each member of the first and second pluralities of images corresponds to a common region of the sample. ### Claim 4 (depends on 1) The method of claim 1, wherein each pixel in the image stack comprises spectral information corresponding to at least 5 different spectral bands of illumination light incident on the sample or emission light from the sample. ### Claim 5 (depends on 1) The method of claim 1, further comprising using the electronic processor to unmix the image stack to obtain one or more component images, each of the component images comprising contributions from substantially only one component in the sample. ### Claim 6 (depends on 5) The method of claim 5, wherein one of the component images corresponds to autofluorescence from the sample. ### Claim 7 (depends on 5) The method of claim 5, wherein one of the component images corresponds to a component of the sample other than autofluorescence. ### Claim 8 (depends on 5) The method of claim 5, further comprising using the electronic processor to analyze the sample based on the one or more component images to determine locations of features within the sample. ### Claim 9 (depends on 1) The method of claim 1, wherein each member of the first and second pluralities of images corresponds to fluorescence emission from the sample. ### Claim 10 (depends on 9) The method of claim 9, wherein each member of the first plurality of images corresponds to fluorescence emission from the sample following illumination of the sample with light in a different wavelength band, and wherein each member of the second plurality of images corresponds to fluorescence emission from the sample following illumination of the sample with light in a different wavelength band. ### Claim 11 (depends on 10) The method of claim 10, further comprising using a color camera to obtain the first and second pluralities of images. ### Claim 12 A system, comprising: an illumination source configured to illuminate a sample with illumination light; a detector configured to obtain one or more images of the sample; and an electronic processor coupled to the detector and configured to: obtain a first plurality of images of the sample, wherein each image in the first plurality of images corresponds to a different spectral band of illumination light incident on the sample from the illumination source or emission light from the sample; obtain a second plurality of images of the sample, wherein each image in the second plurality of images corresponds to a different spectral band of illumination light incident on the sample from the illumination source or emission light from the sample; align the first and second pluralities of images based on information from a first image from the first plurality of images and a second image from the second plurality of images, wherein the first and second images correspond to a shared spectral band; and combine at least some members of the first plurality of images and at least some members of the second plurality of images to form an image stack. ### Claim 13 (depends on 12) The system of claim 12, wherein each pixel in the image stack comprises spectral information derived from at least one member of the first plurality of images and spectral information derived from at least one member of the second plurality of images. ### Claim 14 (depends on 12) The system of claim 12, wherein the electronic processor is configured to align the first and second pluralities of images by spatially registering the first and second pluralities of images so that each member of the first and second pluralities of images corresponds to a common region of the sample. ### Claim 15 (depends on 12) The system of claim 12, wherein each pixel in the image stack comprises spectral information corresponding to at least 5 different spectral bands of illumination light incident on the sample or emission light from the sample. ### Claim 16 (depends on 12) The system of claim 12, wherein the electronic processor is configured to unmix the image stack to obtain one or more component images, each of the component images comprising contributions from substantially only one component in the sample. ### Claim 17 (depends on 16) The system of claim 16, wherein one of the component images corresponds to autofluorescence from the sample. ### Claim 18 (depends on 16) The system of claim 16, wherein one of the component images corresponds to a component of the sample other than autofluorescence. ### Claim 19 (depends on 16) The system of claim 16, wherein the electronic processor is configured to analyze the sample based on the one or more component images to determine locations of features within the sample. ### Claim 20 (depends on 12) The system of claim 12, wherein each member of the first and second pluralities of images corresponds to fluorescence emission from the sample. ### Claim 21 (depends on 20) The system of claim 20, wherein each member of the first plurality of images corresponds to fluorescence emission from the sample following illumination of the sample with light in a different wavelength band, and wherein each member of the second plurality of images corresponds to fluorescence emission from the sample following illumination of the sample with light in a different wavelength band. ### Claim 22 (depends on 21) The system of claim 21, wherein the detector comprises a color camera, and wherein the electronic processor is configured to use the color camera to obtain the first and second pluralities of images.