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How to distinguish necrosis from apoptosis
Rebecca Riggins
Friday, 22 February 2008 23:54 UTC
One of our projects is focused on the mechanisms by which breast cancer cells (the ER-negative MDA-MB-231 cell line) develop resistance to Taxanes. I’d like to be able to differentiate apoptosis from necrosis by immunofluorescence microscopy using a combination of Hoechst and propidium iodide dyes (for example, see the manuscript by McKeague et al., PubMed ID
12556971)
Does anyone out there have experience with this, or any alternate suggestions? Thanks!
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Replies
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When we were looking at cell death in mouse liver (induced by anti-Fas), we found that the only reliable method to distinguish apoptotic from necrotic cells was using morphological criteria (H&E staining). This is a pain, I know…
At the time, most clinical publications were reporting results from TUNEL staining with supporting data from immunohistochemistry for p53, Bcl-2 and/or Bax.
I suppose an alternative now might be immunohistochemistry of a favourite activated caspase… -
Thanks, Bronwen! I really appreciate the suggestion. Pain or not, I’m certainly open to trying H&E to distinguish the two, perhaps coupled with staining for activated caspases or other apoptosis-associated proteins.
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