We consistently isolate intact/functional mitochondria from liver and heart tissue using manual homogenization (to disrupt the cell membrane) followed by differential centrifugation.
I am now trying to disrupt a smaller cell type, and after homogenization (with Potter-Elvehjem and Dounce) the cells remain intact, therefore I cannot isolate the mitochondria.
Does anyone know how to disrupt the cell membrane of small cells without harming the mitochondrial membrane?
Hi Nicole, have you tried incubating your cells in a hypotonic solution such that cells begin to swell and enlarge, and then follow with dounce homogenization? I think there may be a window of salt concentration where the plasma membrane can be sensitized to bursting while mitochondrial membranes remain intact. You may be able to quantify this by titrating progressively lower amounts of salt and assaying cell size by microscopy/coulter counter.
Thanks,
Barry
- Badges
- Science topic
- Similar topics
- Biological Science
- Anatomy
- Tissue