| MITOCHONDRIAL POTENTIAL IMAGING BY FLUORESCENCE AND CONFOCAL MICROSCOPY. |  |
Cells are loaded with JC-1 (1 µg/ml) for 10 min and subjected to confocal microscopy with a BIO-RAD laser scanning system (Radiance 2100) coupled to a Nikon eclipse TE2100U, inverted microscope. For TMRE measurements, cells were loaded with TMRE (100 nM) for 30 min at room temperature, placed on the perfusion chamber of a Zeiss Axiovert 100 TV inverted microscope and superfused continuously with prewarmed (37ºC) standard medium. Fluorescence images are taken at 5 s intervals with a Hamamatsu VIM photon counting camera handled with an Argus-20 image processor.
Human colon carcinoma cells were loaded with TMRE 100 nM for 30 min and subjected to fluorescence microscopy. TMRE accumulated in mitochondria reporting mitochondrial potential. Salicylate induced exit of probe revealing mitochondrial depolarisation. The FCCP uncoupler released all the remaining TMRE revealing colapse of mitochondrial potential. Trace bellow shows averaged TMRE fluorescence of the microscopic field. |
For further details see:
|
