UV-excited autofluorescence responses from cells embedded in turbid media containing hemoglobin analyzed using spectral phasors

Paul Urayama; Mary McNeill; Blake McClain; Nick Majer; Karthik Vishwanath

doi:10.1117/12.2650783

16 March 2023 UV-excited autofluorescence responses from cells embedded in turbid media containing hemoglobin analyzed using spectral phasors

Paul Urayama, Mary McNeill, Blake McClain, Nick Majer, Karthik Vishwanath

Author Affiliations +

Proceedings Volume 12391, Label-free Biomedical Imaging and Sensing (LBIS) 2023; 123910I (2023) https://doi.org/10.1117/12.2650783
Event: SPIE BiOS, 2023, San Francisco, California, United States

Abstract

UV-excited autofluorescence spectroscopy can provide information on the metabolic status of cellular systems, but applications to turbid media such as tissues can be complicated by the presence of multiple scattering, intrinsic absorption, and background fluorescence. Our broader aim is the sensing of cellular-level metabolic status in tissue based the real time assessment of autofluorescence signals using spectral phasor analysis. Previously, we analyzed metabolic responses in yeast cells embedded in turbid media containing significant background fluorescence from collagen. Not only were changes in metabolism detectable under these conditions, but responses associated with NADH- and NADPH-linked metabolisms could also be distinguished. NADH and NADPH are metabolic co-factors having nearly identical excited-state emission but playing significant and distinct roles in cellular metabolism. Here, we extend the phasor analysis approach by sensing metabolic responses of yeast cells embedded in turbid media containing hemoglobin as a source of optical absorption. A metabolic response is induced by chemical perturbation, e.g., by adding cyanide to inhibit cellular respiration or by adding peroxide to induce oxidative stress. We demonstrate that phasor analysis is a versatile tool, e.g., by showing that spectral response associated with changes to cellular metabolism versus optical absorption are spectrally distinct and cannot be accounted for using a two-component spectral model.

Conference Presentation

Citation Download Citation

Paul Urayama, Mary McNeill, Blake McClain, Nick Majer, and Karthik Vishwanath "UV-excited autofluorescence responses from cells embedded in turbid media containing hemoglobin analyzed using spectral phasors", Proc. SPIE 12391, Label-free Biomedical Imaging and Sensing (LBIS) 2023, 123910I (16 March 2023); https://doi.org/10.1117/12.2650783

ACCESS THE FULL ARTICLE

INSTITUTIONAL
Select your institution to access the SPIE Digital Library.

SELECT YOUR INSTITUTION

PERSONAL
Sign in with your SPIE account to access your personal subscriptions or to use specific features such as save to my library, sign up for alerts, save searches, etc.

PERSONAL SIGN IN

No SPIE Account? Create one

PURCHASE THIS CONTENT

SUBSCRIBE TO DIGITAL LIBRARY

50 downloads per 1-year subscription

Members: $195

Non-members: $335 ADD TO CART

25 downloads per 1 - year subscription

Members: $145

Non-members: $250 ADD TO CART

PURCHASE SINGLE ARTICLE

Includes PDF, HTML & Video, when available