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Measuring mitochondrial membrane potential using the FLUOstar Omega microplate reader

Heather Mortiboys, Oliver Bandmann Academic Neurology Unit, University of Sheffield 07/2008

Measurement of the mitochondrial membrane potential is useful in a wide variety of research areas and mitochondrial dysfunction is implicated in diseases such as cancer, diabetes, Parkinson's disease, and stroke.

 

Mitochondrial membrane potential is usually measured using non-invasive cationic dyes. These dyes are sequestered into the mitochondrial matrix in amounts directly proportional to the membrane potential. The dye tetramethylrhodamine methyl ester (TMRM) was used in this study to determine the mitochondrial membrane potential in primary fibroblasts in response to mitochondrial toxins. The fluorescence was related to the cell number determined by the ethidium homodimer fluorophore after cell lysis.

 

These results show that the FLUOstar® Omega is suitable for measuring mitochondrial membrane potential in both 96 and 384 well plate formats. The results are reproducible across a number of different control fibroblast lines and the rescue experiment with an experimental compound proves its suitability for high throughput screening.

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