I am working with stromal cells that have been fixed- I am wondering if it is possible to do hoescht and nile red stains on them like I did with stromal cells in culture
Of course you can.
Hoechst dyes have strong affinity for AT rich chromation regions, rergardless of the living/dead status of the cell. Most commonly used Hoechst dyes are bis-benzimide Hoechst 33258 and 33342, but other variants are also available such as H34580 ...
Nile dyes have strong affinity with lipids, no matter if they are in living cells, dead cells, mayonnaise or yoghurt.
So you can use without any problem those dyes on fixed cells.
Oliv
I would say, it depends on the way, how cells have been fixed. Some fixatives destroy or alter lipid structures, which would result in disturbed lipid staining by Nile Red. If possible, I would suggest labeling with Nile Red before fixation and use a gentle fixation method, such as paraformaldehyde, 2% 10 min on ice.
Yes, it works! But as Daniel suggests, it is better to stain before fixation to avoid artifacts by disrupted membranes.
Good luck :)
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