4.6 Article

Hyperspectral imaging and spectral unmixing for improving whole-body fluorescence cryo-imaging

Journal

BIOMEDICAL OPTICS EXPRESS
Volume 12, Issue 1, Pages 395-408

Publisher

OPTICAL SOC AMER
DOI: 10.1364/BOE.410810

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Funding

  1. National Cancer Institute [R01CA184354, R01CA188491]
  2. Norris Cotton Cancer Center (SYNERGY Translational Pilot Grant)

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By utilizing spectral unmixing, the approach isolates the fluorophore of interest from autofluorescence and other fluorescent reporters, leading to improved detection limits, increased tumor contrast, and significant changes in image interpretation.
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 cryoimaging 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. (C) 2020 Optical Society of America under the terms of the OSA Open Access Publishing Agreement

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