[Study of the relationship between apoptosis and intracellular pH in single living cells using a three-channel real-time fluorescence imaging method].
Using a Dual-View wavelength splitter, a custom-made filter block and only a single intensified charge-coupled device (ICCD), a three-channel real-time fluorescence imaging method for single living cell research was established based on an ICCD fast fluorescence micro-imaging system. The relevant calibrating method for images was also developed to eliminate the influence introduced by spectral crosstalk. Double-labeled with two fluorescent indicators, Annexin V-FITC and SNARF-1, mouse thymocyte was treated with S-nitrosoglutathione (GSNO) and the relationship between their apoptosis and intracellular pH changes was studied in real-time at the single living cell level. The results not only showed the different regularities of intracellular pH changes between apoptosis induced by GSNO and spontaneous apoptosis, but also provided evidence for the application of the three-channel real-time fluorescence imaging method to bio-medical studies.
Lin
Dan-Ying
DY
Key Laboratory for Atomic and Molecular Nanosciences of Education Ministry, Department of Physics, Tsinghua University, Beijing 100084, China. lindy05@mails.thu.edu.cn
Liu
Xiao-Chen
XC
Ma
Wan-Yun
WY
D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D002470:Cell Survival; D006863:Hydrogen-Ion Concentration; D042541:Intracellular Space / Q000737:chemistry* / Q000187:drug effects; D051379:Mice; D008807:Mice, Inbred BALB C; D057054:Molecular Imaging / Q000379:methods*; D026422:S-Nitrosoglutathione / Q000494:pharmacology; D013997:Time Factors
D026422:S-Nitrosoglutathione