Quantitation in immunocytochemistry: correlation of immunogold labeling to absolute number of membrane antigens

Abstract

Baby hamster kidney cells infected with Semliki Forest virus were used as a model system for quantitative immunocytochemical labeling studies. In this system, a well-characterized membrane protein complex is present in different concentrations in three separate locations. Using immunogold labeling of cryosections, we compared the number of gold particles labeling the membranes of endoplasmic reticulum, Golgi stack, and fully formed virions at the plasma membrane to the biochemically determined concentrations. The efficiency of labeling was 40, 13, and 14% for the three structures, respectively. In a comparative study, Lowicryl K4M sections were found to give significantly lower levels of labeling.