to whom it may concern the problem i have is that most people fix yeast cells before adding the hoechst stain to them for DAPI filter images of the nucleus , but i want to stain live yeast cells as fixing cells bleaches the GFP fluorescence which i want to localize to the nuclear region. so i need to get a protocol for hoechst staining of LIVE yeast cells, without fixing them. i have tried adding hoechst to a liquid culture but the cells dont seem to take up the dye unless they are fixed. i would appreciate any suggestions with regard to this problem thanks roopa
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