Analysis of Mitochondrial pH and Ion Concentrations
Vandeven, Martin × Balut, Corina Baron, Szilvia Smets, Ilse Steels, Paul Ameloot, Marcel #
Methods in molecular biology (Clifton, N.J.) vol:591 pages:275-309
Detailed practical information is provided with emphasis on mapping cytosolic and mitochondrial pH, mitochondrial Na(+), and briefly also aspects related to mitochondrial Ca(2+) measurements in living cells, as grown on (un)coated glass coverslips. This chapter lists (laser scanning confocal) microscope instrumentation and setup requirements for proper imaging conditions, cell holders, and an easy-to-use incubator stage. For the daily routine of preparing buffer and calibration solutions, extensive annotated protocols are provided. In addition, detailed measurement and image analysis protocols are given to routinely obtain optimum results with confidence, while avoiding a number of typical pitfalls.