US 12,458,276 B2
Methods for non-invasive, label-free imaging of cellular immune response in human skin using a nonlinear optical microscopy imaging system
Mihaela Balu, Irvine, CA (US); and Anand Ganesan, Irvine, CA (US)
Assigned to The Regents of the University of California, Oakland, CA (US)
Filed by The Regents of the University of California, Oakland, CA (US)
Filed on May 26, 2023, as Appl. No. 18/202,874.
Claims priority of provisional application 63/346,312, filed on May 26, 2022.
Prior Publication US 2023/0380752 A1, Nov. 30, 2023
Int. Cl. A61B 5/00 (2006.01)
CPC A61B 5/414 (2013.01) [A61B 5/0071 (2013.01); A61B 5/444 (2013.01); A61B 5/4848 (2013.01)] 20 Claims
OG exemplary drawing
 
1. A noninvasive method for the in vivo identification and characterization of various immune cell populations in human skin, comprising:
imaging a region of human skin from a subject to detect immune cells noninvasively by using a nonlinear optical imaging system that generates depth-resolved images over large areas with sub-micron resolution based on fluorescence signals generated from one or more endogenous biomolecules found in human skin, wherein one of the endogenous biomolecules is the reduced form of nicotinamide adenine dinucleotide (NADH);
analyzing the image(s) generated from the nonlinear optical imaging system to retrieve information about the metabolic signatures of the detected immune cells by temporal binning the decays of fluorescence lifetime signals of the one or more endogenous biomolecules, and performing slope fluorescence decay analysis; and
distinguishing various immune cell populations based on their morphological and metabolic signatures by using computational analysis.