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Research Papers

Differentiation of apoptosis from necrosis by dynamic changes of reduced nicotinamide adenine dinucleotide fluorescence lifetime in live cells

[+] Author Affiliations
Hsing-Wen Wang, Vladimir Gukassyan

National Yang-Ming University, Institute of Biophotonics, Taipei, Taiwan

Chien-Tsun Chen, Yau-Huei Wei

National Yang-Ming University, Institute of Biochemistry and Molecular Biology, Taipei, Taiwan

Han-Wen Guo, Jia-Sin Yu, Fu-Jen Kao

National Yang-Ming University, Institute of Biophotonics, Taipei, Taiwan

J. Biomed. Opt. 13(5), 054011 (December 04, 2007April 04, 2008April 18, 2008September 10, 2008). doi:10.1117/1.2975831
History: Received December 04, 2007; Revised April 04, 2008; Accepted April 18, 2008; Published September 10, 2008
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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 1.3ns. The NADH average fluorescence lifetime increased to 3.5ns within 15min after 1μM STS treatment and gradually decreased thereafter. The NADH fluorescence intensity increased within 15min. In contrast, no significant dynamic lifetime change was found in cells treated with 1mMH2O2. 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.

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© 2008 Society of Photo-Optical Instrumentation Engineers

Citation

Hsing-Wen Wang ; Chien-Tsun Chen ; Han-Wen Guo ; Jia-Sin Yu ; Yau-Huei Wei, et al.
"Differentiation of apoptosis from necrosis by dynamic changes of reduced nicotinamide adenine dinucleotide fluorescence lifetime in live cells", J. Biomed. Opt. 13(5), 054011 (December 04, 2007April 04, 2008April 18, 2008September 10, 2008). ; http://dx.doi.org/10.1117/1.2975831


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