A fixation method for improved antibody penetration in electron microscopical immunoperoxidase studies.
A fixation method for electron microscopical immunoperoxidase staining has been developed, which (a) allows penetration of antibodies through cell membranes to intracellular antigen sites, (b) provides a reasonable cell preservation and (c) does not alter the antigenic structure in too great an extent. Penetration of the antibodies has been achieved by using saponin as a cell membrane attacking agent. The best results could be obtained after pretreatment of cell monolayers with a mixture of 0.05% saponin, 0.0125%-0.05% glutaraldehyde and 1% paraformaldehyde for 5 min at 4 degrees C, and postfixing them with the corresponding fixative without saponin for 45 min at 4 degrees C.
D000906:Antibodies; D000941:Antigens / Q000032:analysis*; D002460:Cell Line; D007124:Immunoenzyme Techniques; D007202:Indicators and Reagents; D008854:Microscopy, Electron; D013194:Staining and Labeling
D000906:Antibodies; D000941:Antigens; D007202:Indicators and Reagents