PMID- 19647441
OWN - NLM
STAT- MEDLINE
DCOM- 20091102
LR  - 20090904
IS  - 1878-4291 (Electronic)
IS  - 0968-4328 (Linking)
VI  - 40
IP  - 8
DP  - 2009 Dec
TI  - Quantitative analysis of caspase-3 activation by fitting fluorescence emission 
      spectra in living cells.
PG  - 811-20
LID - 10.1016/j.micron.2009.07.001 [doi]
AB  - Confocal fluorescence imaging and fluorescence resonance energy transfer (FRET) 
      technology have been widely used to study protein-protein interactions in living 
      cells. However, it is very difficult to quantitatively analyze FRET efficiency 
      due to the excitation spectral crosstalk and emission spectral crosstalk between 
      donor and acceptor. In this study, we developed a novel method to quantitatively 
      obtain the FRET efficiency by fitting the emission spectra (FES) of 
      donor-acceptor pair, and this method is free from both excitation and emission 
      spectral crosstalk. We used the FES method to quantitatively monitor the FRET 
      efficiency of SCAT3, a caspase-3 indicator based on FRET, inside living cells 
      stably expressing SCAT3 during STS-induced apoptosis. At 0, 6 and 12 h after STS 
      treatment, the FRET efficiency of SCAT3 obtained by FES are consistent with that 
      by two-photon excitation (TPE) fluorescence lifetime imaging microscopy (FLIM) in 
      living cells stably expressing SCAT3. In this study, the FES was also used to 
      analyze the caspase-3 activation in living cells during anti-cancer drug such as 
      taxol, Artesunate (ART) or Dihydroartemisinin (DHA) treatment. Our results showed 
      that ART or DHA induced apoptosis by a caspase-3-dependent manner, while 
      caspase-3 was not involved in taxol-induced cell death.
FAU - Wang, Longxiang
AU  - Wang L
AD  - MOE Key laboratory of Laser Life Science & Institute of Laser Life Science, South 
      China Normal University, Guangzhou 510631, China.
FAU - Chen, Tongsheng
AU  - Chen T
FAU - Qu, Junle
AU  - Qu J
FAU - Wei, Xunbin
AU  - Wei X
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20090723
PL  - England
TA  - Micron
JT  - Micron (Oxford, England : 1993)
JID - 9312850
RN  - 0 (Antineoplastic Agents)
RN  - EC 3.4.22.- (Caspase 3)
SB  - IM
MH  - Antineoplastic Agents/pharmacology
MH  - Caspase 3/*analysis
MH  - Cell Line, Tumor
MH  - Cell Survival
MH  - Epithelial Cells/drug effects
MH  - Fluorescence Resonance Energy Transfer/*methods
MH  - Humans
MH  - Image Processing, Computer-Assisted/*methods
EDAT- 2009/08/04 09:00
MHDA- 2009/11/03 06:00
CRDT- 2009/08/04 09:00
PHST- 2009/03/23 00:00 [received]
PHST- 2009/06/24 00:00 [revised]
PHST- 2009/07/02 00:00 [accepted]
PHST- 2009/08/04 09:00 [entrez]
PHST- 2009/08/04 09:00 [pubmed]
PHST- 2009/11/03 06:00 [medline]
AID - S0968-4328(09)00110-3 [pii]
AID - 10.1016/j.micron.2009.07.001 [doi]
PST - ppublish
SO  - Micron. 2009 Dec;40(8):811-20. doi: 10.1016/j.micron.2009.07.001. Epub 2009 Jul 
      23.