I have been dual staining cells treated with drugs and at lower doses I get the characteristic shift to Hoechst stained and then dual stained with hoechst and 7-AAD stained. However for a higher dosage of the drug the dot plot showed a shift in the 7-AAD staining without hoechst. Has anyone else found this? Or offer any explanation as to why this would occur?
Many thanks in advance
When you use these markers occur this:
Hoechst is used for apoptosis, and 7AAD: used to indicate a disruption in membrane. Maybe in your lower doses you have this disruption. In higher doses the membrane was completely ruptured and you can't mark the nucleus with Hoechst.
When you use these markers occur this:
Hoechst is used for apoptosis, and 7AAD: used to indicate a disruption in membrane. Maybe in your lower doses you have this disruption. In higher doses the membrane was completely ruptured and you can't mark the nucleus with Hoechst.
I agree with Cássio Prinholato da Silva
Hoechst: Chromatin condensation (apoptosis)
7AAD: membrane integrity (viability)
You cells: faster apoptosis or necrosis
Using Ethidium and Propidium you can see that dead cell show PI fluorescence only as it displaces the EB. With Hoechst and PI you can get energy transfer between the two.
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