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

作者：Wang Hsing Wen^*; Gukassyan Vladimir; Chen Chien Tsun; Wei Yau Huei; Guo Han Wen; Yu Jia Sin; Kao Fu Jen

来源：Journal of Biomedical Optics, 2008, 13(5): 054011.

DOI：10.1117/1.2975831

• nicotinamide adenine dinucleotide intrinsic florescence
• two-photon fluorescence lifetime imaging
• apoptosis detection
• noninvasive optic probe
• treatment outcome

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 (H(2)O(2))-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 similar to 1.3 ns. The NADH average fluorescence lifetime increased to similar to 3.5 ns within 15 min after 1 mu M 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 H(2)O(2). 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.

• 出版日期2008-10