Immunofluorescent staining of pancreatic sections v1

This protocol describes the steps for performing fluorescence immunohistochemistry on fixed-frozen pancreatic tissue sections. It is suitable for pancreatic tissue isolated from rats and mice at postnatal to adult stages. We routinely apply this protocol to quantify total and proliferating or apoptotic beta-, alpha- and delta-cells. Briefly, pancreata are fixed in 4% paraformaldehyde solution and cryoprotected overnight in 30% sucrose. Tissue are then embedded, frozen, sectioned and mounted on slides. Antigen retrieval is performed using sodium citrate buffer prior to immunostaining.