I have 4T1-Luc cells seeded in MatTek 6-well plates, 10^5 cells/well. After I treat them with a nanoparticle-dye conjugate I want to fix them for imaging. What is the best procedure for this? I am a chemist so I'm hoping someone with cell culture experience can point me in the right direction.
Dear Colin,
Depending on the cell line and on the assay to do with them, we genreally apply different Fixation protocols. So I can't tell you the right answer to that question, but I can make some suggestions what you could try.
1) Fixation in 4% PFA/PBS. Wash your cells with PBS, then add 4%PFA/PBS and incubate @RT for 10-15min. Then wash again and store your cells in PBS for short term or 1%PFA for long-term.
2) Use cytofix/cytoperm from BD Biosciences according to the manufacturer's recommendations. However, if you don't want to permeabilize your cells, that might not be the best option.
3) Fixation in ice-cold methanol. Wash your cells with PBS, then add ice-cold methanol and incubate on ice for 10min. Then wash again and store your cells in PBS for short term or 1%PFA for long-term.
I hope this helps somehow.
Good luck,
Cindy
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