Caco-2 cells were grown on cover slips and stained with Acridine orange and Ethidium bromide to show apoptosis, All cells show orange color , I have tried acridine orange alone it show all cells green under blue fluorescence, I did not fix cells . please I need help or clear protocol for staining technique to show apoptosis in my cell line with the right colors (orange, green) of live and apoptotic cells , as I don’t have possibility to use other methods.
Dear Hebat,
Acridine orange is not purposed to evidence apoptosis.
The attached article (Mijatovic et al., 2006) could be of help for the use of acridine orange to study some cell death types other than apoptosis.
Acridine orange is often used to evidence autophagy, but this is non specific.
Acridine orange can emit orange versus green colors dependening on the intracellular ph (see Mijatovic et al., 2006).
Here attached are general articles about how performing various types of assays to evidence apoptosis versus non-apoptotic cell death types.
Best regards
Robert
Dear Hebat,
Acridine orange is not purposed to evidence apoptosis.
The attached article (Mijatovic et al., 2006) could be of help for the use of acridine orange to study some cell death types other than apoptosis.
Acridine orange is often used to evidence autophagy, but this is non specific.
Acridine orange can emit orange versus green colors dependening on the intracellular ph (see Mijatovic et al., 2006).
Here attached are general articles about how performing various types of assays to evidence apoptosis versus non-apoptotic cell death types.
Best regards
Robert
Hello Hebat,
You can try using Annexin V-FITC kit which allows fluorescent detection of annexin V that binds to apoptotic cells . The kit also comes with propidium iodide (PI) that helps in distinguishing the necrotic cells as well from the apoptotic cells.
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