I fixed my cells with 10% formalin and conserved in PBS at 4ºC. We had some problems with the antibodies and I have to put off the IF. For how long can I conserve the formalin-fixed cells?
Hi amaya,
Ideally, you can fix your cells in acetone/ethanol/para-formaldehyde/neutral buffered formalin etc., for 10 minutes (unless you have tissue section that are embedded in paraffin/resin etc., Please note fixing cells in 10% formalin may damage the cells as well as the epitopes/antigen of interest. After fixing the cells, you can wash your cells in PBS for 3-5 minutes and keep repeating this step when you treat cells with primary antibody, secondary antibody etc., 4 degree centigrade is fine for immunostaining methods.
Hope this helps.
Polly
I personally kept them up to 48 hours in 4 degree after fixation and they were OK. I fixed them with PFA and washed them with PBS-1% BSA then immersed them in PBS for 48 hours before IF. I am not sure how far we can go after fixation, that is a good question. Did somebody tried keeping them longer in fridge?
we keep tissue sections that were PFA fixed , in PBS for months in 4degree centigrade, and later do IF as well as IHC . Antigen retrieval becomes a must in those cases.
I think that the best scenario is to keep your cells in formalin for 10 mins, proceed to the permeating step (10 mins.) and then the washing step with the 1X PBS( three times, each time is 10 mins). When you finish all these steps, you can preserve your cells as long as 24-48 hours.
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