US 11,835,463 B2
Multispectral sample imaging
Peter J. Miller, Cambridge, MA (US); Kent S. Johnson, Cambridge, MA (US); and Carla Coltharp, Waltham, MA (US)
Assigned to Akoya Biosciences, Inc., Menlo Park, CA (US)
Filed by Akoya Biosciences, Inc., Menlo Park, CA (US)
Filed on Apr. 14, 2022, as Appl. No. 17/720,751.
Application 17/720,751 is a continuation of application No. 16/403,401, filed on May 3, 2019, granted, now 11,307,142.
Claims priority of provisional application 62/666,697, filed on May 3, 2018.
Prior Publication US 2022/0244183 A1, Aug. 4, 2022
Int. Cl. G01N 21/64 (2006.01); G06T 7/194 (2017.01); G06T 7/00 (2017.01); G01J 3/28 (2006.01)
CPC G01N 21/6458 (2013.01) [G01J 3/28 (2013.01); G01J 3/2823 (2013.01); G01N 21/6428 (2013.01); G06T 7/0012 (2013.01); G06T 7/194 (2017.01); G01J 2003/2826 (2013.01); G01N 2021/6419 (2013.01); G01N 2021/6421 (2013.01); G06T 2207/10036 (2013.01); G06T 2207/10056 (2013.01); G06T 2207/10064 (2013.01); G06T 2207/30024 (2013.01)] 24 Claims
OG exemplary drawing
 
1. A method, comprising:
obtaining one or more images of a biological sample, wherein each sample image corresponds to a different combination of illumination light wavelengths and light emission wavelengths; and
obtaining a background image of the biological sample by exposing the biological sample to background excitation light and measuring light emission from the biological sample at one or more background emission wavelengths that are each within a wavelength range defined by the light emission wavelengths,
wherein for each of one or more spectral contributors in the biological sample exposed to the background excitation light, a spectral emission intensity at each of the background emission wavelengths is 10% or less of a maximum spectral emission intensity of the spectral contributor following excitation at each of the illumination light wavelengths; and
wherein a distribution of the background emission wavelengths has a full width at half maximum (FWHM) spectral width Δλ and center wavelength λc, and the background image corresponds to measured light emission for at least some wavelengths within a range from λc-λ/2 to λc+λ/2.