What is the best method for preparing gelatin methacrylate embedded cells for Immunofluorescence? I have fixed with 4% PFA, embedded in histogel, paraffin embedded and then performed antigen retrieval using citrate buffer at 37 degrees celsius. My antibody fluorescence is very poor/faint. I know gelatin is temperature sensitive so I am hesitant to raise the citrate buffer temperature during retreival and risk my sample falling off the slide. Does anyone have a method that works well for them?

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