Abstract
Staining of intracellular antigens by immunological reagents is dependent on free diffusion of antibodies into and out of cells. This is accomplished by disruption of the cell membrane by organic solvents or detergents. Organic solvents dehydrate the cell and denature proteins, with the effect of preserving the integrity of the cell to varying degrees dependent on the solvent used. The use of detergents is more variable in effect, but does not preserve the cell unless a fixative such as formaldehyde is used. Immunological staining has been employed for many years and the literature documents that different antigen/antibody combinations require different cell pretreatments. Quantitative analysis is relatively new but the same in every respect except that reagents for staining must be used under saturating conditions and controls must be carefully designed. This article is a review of current literature, methods under development in my laboratory, and relevant issues.
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