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Journal of Biomedical Optics

Differentiation of apoptosis from necrosis by dynamic changes of reduced nicotinamide adenine dinucleotide fluorescence lifetime in live cells
Author(s): Hsing-Wen Wang; Vladimir Ghukasyan; Chien-Tsun Chen; Yau-Huei Wei; Han-Wen Guo; Jia-Sin Yu; Fu-Jen Kao
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Paper Abstract

Direct monitoring of cell death (i.e., apoptosis and necrosis) during or shortly after treatment is desirable in all cancer therapies to determine the outcome. Further differentiation of apoptosis from necrosis is crucial to optimize apoptosis-favored treatment protocols. We investigated the potential modality of using tissue intrinsic fluorescence chromophore, reduced nicotinamide adenine dinucleotide (NADH), for cell death detection. We imaged the fluorescence lifetime changes of NADH before and after staurosporine (STS)-induced mitochondria-mediated apoptosis and hydrogen peroxide (H2O2)-induced necrosis, respectively, using two-photon fluorescence lifetime imaging in live HeLa cells and 143B osteosarcoma. Time-lapsed lifetime images were acquired at the same site of cells. In untreated cells, the average lifetime of NADH fluorescence was ~1.3 ns. The NADH average fluorescence lifetime increased to ~3.5 ns within 15 min after 1 μM STS treatment and gradually decreased thereafter. The NADH fluorescence intensity increased within 15 min. In contrast, no significant dynamic lifetime change was found in cells treated with 1 mM H2O2. Our findings suggest that monitoring the NADH fluorescence lifetime may be a valuable noninvasive tool to detect apoptosis and distinguish apoptosis from necrosis for the optimization of apoptosis-favored treatment protocols and other clinical applications.

Paper Details

Date Published: 1 September 2008
PDF: 9 pages
J. Biomed. Opt. 13(5) 054011 doi: 10.1117/1.2975831
Published in: Journal of Biomedical Optics Volume 13, Issue 5
Show Author Affiliations
Hsing-Wen Wang, National Yang-Ming Univ. (Taiwan)
Vladimir Ghukasyan, National Yang-Ming Univ. (Taiwan)
Chien-Tsun Chen, National Yang-Ming Univ. (Taiwan)
Yau-Huei Wei, National Yang-Ming Univ. (Taiwan)
Han-Wen Guo, National Yang-Ming Univ. (Taiwan)
Jia-Sin Yu, National Yang-Ming Univ. (Taiwan)
Fu-Jen Kao, National Yang-Ming Univ. (Taiwan)


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