Real-time single cell analysis of molecular mechanism of apoptosis and proliferation using FRET technique

Tongsheng Chen; Da Xing; Xuejuan Gao; Fang Wang

doi:10.1117/12.710925

27 October 2006 Real-time single cell analysis of molecular mechanism of apoptosis and proliferation using FRET technique

Tongsheng Chen, Da Xing, Xuejuan Gao, Fang Wang

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Proceedings Volume 6047, Fourth International Conference on Photonics and Imaging in Biology and Medicine; 604736 (2006) https://doi.org/10.1117/12.710925
Event: Fourth International Conference on Photonics and Imaging in Biology and Medicine, 2005, Tianjin, China

Abstract

Bcl-2 family proteins (such as Bid and Bak/Bax) and 14-3-3 proteins play a key role in the mitochondria-mediated cell apoptosis induced by cell death factors such as TNF-α and lower power laser irradiation (LPLI). In this report, fluorescence resonance energy transfer (FRET) has been used to study the molecular mechanism of apoptosis in living cells on a fluorescence scanning confocal microscope. Based on the genetic code technique and the green fluorescent proteins (GFPs), single-cell dynamic analysis of caspase3 activation, caspase8 activation, and PKCs activation are performed during apoptosis induced by laser irradiation in real-time. To investigate the cellular effect and mechanism of laser irradiation, human lung adenocarcinoma cells (ASTC-a-1) transfected with plasmid SCAT3 (pSCAT3)/ CKAR FRET reporter, were irradiated and monitored noninvasively with both FRET imaging. Our results show that high fluence lower power laser irradiation (HFLPLI) can induce an increase of caspase3 activation and a decrease of PKCs activation, and that LPLI induces the ASTC-a-1 cell proliferation by specifically activating PKCs.

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Tongsheng Chen, Da Xing, Xuejuan Gao, and Fang Wang "Real-time single cell analysis of molecular mechanism of apoptosis and proliferation using FRET technique", Proc. SPIE 6047, Fourth International Conference on Photonics and Imaging in Biology and Medicine, 604736 (27 October 2006); https://doi.org/10.1117/12.710925

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KEYWORDS

Cell death

Fluorescence resonance energy transfer

Laser irradiation

Microscopes

Luminescence

Molecular mechanisms

Green fluorescent protein

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