Open AccessHyperspectral imaging and spectral unmixing for improving whole-body fluorescence cryo-imaging
Author(s) -
Dennis J. Wirth,
Brook K. Byrd,
Boyu Meng,
Rendall R. Strawbridge,
Kimberley S. Samkoe,
Scott C. Davis
Publication year - 2020
Publication title -
biomedical optics express
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.362
H-Index - 86
ISSN - 2156-7085
DOI - 10.1364/boe.410810
Subject(s) - hyperspectral imaging , autofluorescence , fluorescence lifetime imaging microscopy , fluorophore , fluorescence , spectral imaging , molecular imaging , biodistribution , multispectral image , visualization , computer science , optics , artificial intelligence , chemistry , physics , in vivo , biology , biochemistry , microbiology and biotechnology , in vitro
Whole-animal fluorescence cryo-imaging is an established technique that enables visualization of the biodistribution of labeled drugs, contrast agents, functional reporters and cells in detail. However, many tissues produce endogenous autofluorescence, which can confound interpretation of the cryo-imaging volumes. We describe a multi-channel, hyperspectral cryo-imaging system that acquires densely-sampled spectra at each pixel in the 3-dimensional stack. This information enables the use of spectral unmixing to isolate the fluorophore-of-interest from autofluorescence and/or other fluorescent reporters. In phantoms and a glioma xenograft model, we show that the approach improves detection limits, increases tumor contrast, and can dramatically alter image interpretation.