I'm working with cytospinned human cardiomyocytes and would like to stain several slides all at the same time. My current protocol makes use of 4% PFA with a permeabilization step combined with the blocking step. We observed a maintained membrane integrity with this protocol so we stuck with it. If I want to store the slides long term, can I keep the fixed slides (washed with PBS?) in lower temps? Or would it be necessary to use an organic solvent for this purpose? Thanks
Hi Julius,
Storing your fixed cells in PBS plus 0.02% NaN3 at 4C will suffice. You should also seal the containers to avoid evaporation. I've kept cells for over year like that without any apparent problem.
Cheers
Thank you for the suggestions, Dr Sebastián Dupraz and Ms Rachel Gad
!But just to clarify, these cells are on glass slides right? Does it matter then if i submerge them completely in the PBS supplemented with sodium azide or it's fine pipetting a few microliters on the slide and putting them in a damp chamber? Although I was hoping to store them for like 6 weeks maximum.
Hi Julius,
I keep the coverslips with the attached cells submerged in PBS-NaN3 in the same petri dish were I fixed them. You could use a damp chamber if it is more handy for you but they have higher chances of drying out.
Cheers,
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