Application of Nanosecond Pulsed Electric Fields into HeLa Cells Expressing Enhanced Green Fluorescent Protein and Fluorescence Lifetime Microscopy

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• 作者：Kamlesh Awasthi ; Takakazu Nakabayashi ; Nobuhiro Ohta
• 刊名：The Journal of Physical Chemistry B
• 出版年：2012
• 出版时间：September 13, 2012
• 年：2012
• 卷：116
• 期：36
• 页码：11159-11165
• 全文大小：330K
• 年卷期：v.116,no.36(September 13, 2012)
• ISSN：1520-5207

文摘

An electrode microchamber has been constructed for applying nanosecond pulsed strong electric fields to living cells, and fluorescence lifetime microscopy (FLIM) has been used to investigate the effects of external electric fields on dynamics and function of HeLa cells expressing enhanced green fluorescent protein (EGFP). Both morphological change in cells and reduction of the fluorescence lifetime of EGFP have been observed after application of electric fields having a pulsed width of 50 ns and a strength of 4 MV m^鈥?, indicating that apoptosis, which is a programmed cell death, was induced by nanosecond pulsed electric fields and that fluorescence lifetime of EGFP decreased along with the induction of apoptosis. The reduction of the fluorescence lifetime occurred before the morphological change, indicating that FLIM provides a sensitive and noninvasive detection of the progress of apoptosis induced by application of nanosecond pulsed electric fields.