Flow cytometry in the study of mitochondrial respiratory chain disorders.
We have developed a flow cytometric assay to measure the oxidative capacity of cultured lymphoblasts as a possible screening test for patients suspected of having a defect of the mitochondrial respiratory chain. Cells were incubated overnight in serum free media, followed by incubation with dihydroethidium with and without rotenone, and then analysed using flow cytometry to measure fluorescence. Inhibition with rotenone gave an increase in fluorescence compared to uninhibited cells. The change in fluorescence was significantly lower in four of the six patient cell lines, with a correlation between the activity of complex I and change in fluorescence. This method may be applicable to cell lines with defects in other complexes of the respiratory chain.
Setterfield
Karen
K
Department of Biological Sciences, Macquarie University, North Ryde, Sydney, NSW, Australia.
Williams
Andrew J
AJ
Donald
Jennifer
J
Thorburn
David R
DR
Kirby
Denise M
DM
Trounce
Ian
I
Christodoulou
John
J