1. The cell membrane surface of the target can be fixed before staining (mainly according to the experimental conditions to determine), the target point in the cell must be fixed and then stained;
2.the immune fluorescence experiment is generally used more than 4% formaldehyde fixed 15 minutes, fixed time is not more than 20 minutes (fixed time will affect the protein immunogenicity, and immune fluorescence generally do not do antigen repair). Permeablization:
In order to be free and in the cells, the cells need to be fixed before staining. If it is not fixed, the cells will be cracked.