Organoids have complex 3-dimensional structures and are embedded in ECM hydrogels making characterization difficult. Immunofluorescent (IF) staining of organoids with antibodies can be performed to visualize molecular markers of cellular behavior, proliferation, differentiation, cell health and identity. Whole-mount immunostaining is intended for small pieces of tissue without sectioning and methods are very similar to immunocytochemistry (ICC) or immunohistochemistry (IHC) staining of cryosections. Whole-mount staining of organoids can be used for antibody based characterization. In this organoid staining protocol we detail methods to fix, permeabilize, and stain whole-mount organoids for analysis by immunofluorescent confocal microscopy.
Figure 1.Immunofluorescence of organoids using antibodies. Human epithelial intestinal and lung organoids stained with antibodies for Acetyl-α-Tubulin (A), α-Carbonic Anhydrase IV (B) and Sox-9 (AB5535) (C).