Analysis of cytosolic pH changes in thymocytes during early apoptosis with improved three-channel real-time fluorescence imaging.

Cytosolic pH changes in single thymocytes in the early stages of apoptosis were examined with spectral-crosstalk-improved three-channel real-time fluorescence imaging. Spectral crosstalk was greatly reduced by this improved image processing method. Compared with normal thymocytes, the improved image processing method showed that the cytosolic pH value of thymocytes undergoing apoptosis was lower. A slight diversity in the intracellular pH values of samples subjected to different drug treatments was also observed. This new image processing method is beneficial to the application of the three-channel real-time fluorescence imaging to biological processes.