Evaluation of some cell death features by real time real space microscopy

In the past few years, the investigation of many cell death features, especially ones associated with changes in DeltaPsi(m), have gained an important insights with the development of high-resolution fluorescence microscopy. With the use of real time real space measurements, it was possible to perform dynamic studies not only to investigate the location of the organelles, but also to follow changes in transport mechanism, such as Ca(2+) concentration in different subcellular compartments. In addition, this technique has been used for the simultaneous tracking of organelle location, ion measurements, and DeltaPsi(m,) which clearly contributed to further understanding mechanisms related to the control of cell death. This chapter describes the methodology employed to study changes in DeltaPsi(m), Bax translocation, and Ca(2+) measurements upon apoptotic induction. It also details the new technique developed and employed in our laboratory to measure Ca(2+) signaling in brain slices by confocal microscopy. This method has been applied to investigate real time real space studies in different models of neurodegenerative processes, such as Huntington's disease and aging.