I understand that the number of mitochondria in a cell is difficult to quantify.
1. Is measuring the mtDNA content same as measuring the number of mitochondria?
2. If I opt for the measurement of the mitochondrial mass, is MitoTracker fluorescent probe the best way to go with?
Thank you
1.) Mitochondria are dynamic organelles. In a healthy cell they constantly fuse and divide. Therefore it is not feasible to determine mitochondrial numbers. On the other hand, estimating mitochondrial mass by measuring mtDNA content seems to be a viable option (i. e., measuring the mtDNA/nDNA ratio by qPCR of ND1 (mtDNA) and 18S RNA (nDNA)).
2.) In my opinion it is better to do either the mtDNA measurement or immunodetection of a mitochondrial protein (e. g., VDAC or COX2). Fluorescent markers are often not able to stain all mitochondria in a cell homogenously. Therefore variations in intensity can be very high.
In the attachment you will find an interesting paper in which the aforementioned methods have been applied.
Cheers, Christian
Article Bioenergetic Flux, Mitochondrial Mass, and Mitochondrial Mor...
Hello,
Graciela Rabadan, Please, do you mean by electronic microscopy doing a simple count of mitochondria on an ultra-fine section using a software?
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