Immunoelectron microscopic labeling of immunoglobulin in plasma cells after osmium fixation and epoxy embedding.
Previous studies have found that immunoglobulin cannot be immunolabeled in tissues prepared for electron microscopy by usual methods. To test this conclusion, we used a protein A-gold postembedding immunolabeling method on tissues that were fixed in glutaraldehyde, post-fixed in osmium tetroxide, and embedded in epoxy resin; sections were pretreated with sodium metaperiodate. A variety of common fixation protocols were also used and the most suitable conditions for immunolabeling were determined. This technique permitted the ultrastructural localization of immunoglobulin light chains in optimally preserved and contrasted plasma cells from human tonsil, lymph nodes, plasmacytomas, and a renal biopsy. We were able to demonstrate multiple antigens in the same tissue and label antigens in tissues that had been stored for many years in epoxy resin. The technique allows quantitation of the gold label over plasma cell organelles and therefore gives information about the immunoglobulin secretory pathway in these cells. We found that the protein A-gold procedure compares favorably in technical ease with the immunoperoxidase, avidin-biotin peroxidase, and immunoglobulin-colloidal gold immunolabeling methods, and has added advantages in allowing precise localization and quantitation of the labeled antigen.
D004852:Epoxy Compounds; D005976:Glutaral; D006652:Histological Techniques; D006801:Humans; D008198:Lymph Nodes / Q000648:ultrastructure; D008854:Microscopy, Electron; D009993:Osmium Tetroxide; D014066:Palatine Tonsil / Q000648:ultrastructure; D010950:Plasma Cells / Q000648:ultrastructure*
D004852:Epoxy Compounds; D009993:Osmium Tetroxide; D005976:Glutaral