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Proceedings Paper

Spectral phasor analysis of autofluorescence responses from cells embedded in turbid media containing collagen
Author(s): Max Kreider; Andrew I. Rodriguez; Karthik Vishwanath; Paul Urayama
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Paper Abstract

Autofluorescence spectroscopy can provide information on the metabolic status of cellular systems, but extensions of these techniques to turbid media such as tissues is complicated by the presence of multiple scattering, background fluorescence, and intrinsic absorption. Phasor analysis is a class of analytical approaches for the real-time assessment of emission signals that could be used to decipher cellular-level metabolic status of tissues. Spectral phasor analysis was originally developed for the rapid segmentation of hyperspectral images and has since been used for monitoring cellular NAD(P)H conformation from UV-excited cellular autofluorescence. Specifically, we showed previously that chemically induced autofluorescence responses in Saccharomyces cerevisiae (baker’s yeast) suspensions could not be accounted for using the two-component free vs. protein-bound model for conformation. Rather, by considering a series of physically similar and dissimilar chemicals acting on multiple metabolic pathways, we showed that responses affecting different pathways, e.g., involving cellular respiration versus oxidative stress, could be distinguished. Here, we seek to extend this pathway-level interpretation to the sensing of cellular metabolism in tissues by monitoring the cyanide-induced metabolic response of yeast cells embedded in media containing 9-cyanoanthracene or collagen as sources of background emission. Despite the similarity between autofluorescence and background spectra, we observe spectral behavior consistent with the discrimination of the metabolic response from the background emission. Performance over specifically selected noncontinuous spectral bands to reject chromophore absorption is also assessed.

Paper Details

Date Published: 20 February 2020
PDF: 10 pages
Proc. SPIE 11251, Label-free Biomedical Imaging and Sensing (LBIS) 2020, 112510B (20 February 2020); doi: 10.1117/12.2546398
Show Author Affiliations
Max Kreider, Miami Univ. (United States)
Andrew I. Rodriguez, Miami Univ. (United States)
Karthik Vishwanath, Miami Univ. (United States)
Paul Urayama, Miami Univ. (United States)

Published in SPIE Proceedings Vol. 11251:
Label-free Biomedical Imaging and Sensing (LBIS) 2020
Natan T. Shaked; Oliver Hayden, Editor(s)

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