Analysis of mitochondrial pH and ion concentrations.
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.
vandeVen
Martin
M
Cell Physiology Group, Biomedical Research Institute, Hasselt University, Diepenbeek, Belgium.
Balut
Corina
C
Baron
Szilvia
S
Smets
Ilse
I
Steels
Paul
P
Ameloot
Marcel
M
D000818:Animals; D002118:Calcium / Q000378:metabolism*; D002138:Calibration; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism; D002470:Cell Survival; D003600:Cytosol / Q000378:metabolism; D004285:Dogs; D005456:Fluorescent Dyes / Q000378:metabolism; D005898:Glass; D006863:Hydrogen-Ion Concentration; D007202:Indicators and Reagents / Q000378:metabolism; D018613:Microscopy, Confocal; D008928:Mitochondria / Q000378:metabolism*; D057054:Molecular Imaging; D009068:Movement; D012964:Sodium / Q000378:metabolism*; D012984:Software
D005456:Fluorescent Dyes; D007202:Indicators and Reagents; D012964:Sodium; D002118:Calcium