Immunofluorescence of colon issues.
Paraformaldehyde-fixed colon tissues were embedded in paraffin for analysis of F4/80+ cell infiltration. Sections were washed three times with PBS and then exposed to 3.0% H2O2 for 1h to block endogenous peroxidase activity. Subsequently, the sections were blocked with 3% BSA for 30min at room temperature and then incubated at 4°C overnight with anti-F4/80 (#GB11027, 1:500 per mouse, Servicebio). The next day, sections were placed in PBS (PH7.4) and washed 3 times on a decolorizing shaker for 5min each time. The corresponding secondary antibody was added and incubated for 50min at room temperature and away from light. Fluorescence microscopy to analyze the results (Zeiss Axioplan 2).