Immunofluorescence microscopy is a powerful method for analysis of the subcellular localization of the protein of interest. The use of fluorescence is very effective for multiple labeling and for higher magnification observation with a laser confocal microscope as well as a conventional fluorescence microscope. The technique is easy and generally used for researchers; however, it is also true that they sometimes show the results inappropriately and incorrectly. I would like to introduce the basic methods with some helpful suggestions of the immunofluorescent staining in cultured cells, especially focusing on the followings:
1) Cell culture for immunocytochemistry
2) Fixation
3) Treatment before immunostaining
4) Key points for multiple labeling: choices of the secondary antibodies and fluorescence dyes
5) Observation and storage of the samples
I hope this talk could help researchers in the Pharmacological Society to get better and beautiful results in immunofluorescence microscopy.