Hoechst Stain does it stain dead cells - (Nov/02/2010 )

Hello.

I'm using the Hoechst Stain to determine cell proliferation. Ideally I would only like to include the live cells so does the Hoechst stain distinguish between dead and live cells. Based on how it works I imagine it doesn't but as I am working with adherent cells which are washed before I do the stain would I be correct in saying that the majority of dead cells will have been washed away?

Any advice would be greatly appreciated

Hoechst stains all cells if you fix them. I haven't tried staining unfixed cells though.

Hi,

there are different Hoechst stains available: Hoechst 33342 and 33258 are definitly cell permeable, so it will stain all cells if dead or alive even without fixation. I use 5µl Hoechst <2mg/ml stock> per ml staining solution (Media or PBS) and incubate living cells for 10 min. at 37°C for microscopy.

great thanks for the replies

Just wondering would ye consider for presenting results on proliferation that a stain such as Hoeschst is acceptable or should it be used in conjunction with a cell viability assay? I've found a number of papers that dont seem to use to use any viability assay.