Immunoelectron microscopic (IEM) studies on glutaraldehyde-fixed renal specimen.
Immunofluorescent microscopy (IF) has become an essential tool in the routine diagnosis of renal pathology. It is thought that glomerular IF patterns represent different forms of immunologically mediated glomerular lesions. However, in order to preserve antigenicity IF is usually done on frozen, unfixed specimen by light microscopy and it is difficult to locate target macromolecules in tissue with precision. In order to establish the precise location of such macromolecules in glomeruli in relation to the ultrastructure, we undertook an immunoelectron microscopic (IEM) study on renal biopsies. Percutaneously biopsied tiny specimen was fixed in glutaraldehyde and processed with protease. Using peroxidase-labeled antisera, a direct IEM was done. With this technique, target macromolecules corresponded well not only to electron-dense deposits but also to IF patterns. In one case of membranous nephropathy, immunoglobulin (Ig) was found to be outside of the glomerular basement membrane (GBM), while IgA was seen to be inside the GBM. For this same case, a similar localization of immunoglobulins was not revealed by IF. In conclusion, IEM is a useful technique that can broaden our knowledge of pathogenic mechanisms in glomerular disease.
D000328:Adult; D002648:Child; D003176:Complement C3 / Q000032:analysis; D005260:Female; D005404:Fixatives; D005921:Glomerulonephritis / Q000276:immunology; D005976:Glutaral; D006651:Histocytochemistry; D006735:Horseradish Peroxidase; D006801:Humans; D007070:Immunoglobulin A / Q000032:analysis; D007074:Immunoglobulin G / Q000032:analysis; D007075:Immunoglobulin M / Q000032:analysis; D007158:Immunologic Techniques; D007678:Kidney Glomerulus / Q000276:immunology* / Q000648:ultrastructure; D008297:Male; D008854:Microscopy, Electron / Q000379:methods*; D010447:Peptide Hydrolases / Q000378:metabolism; D013048:Specimen Handling
D003176:Complement C3; D005404:Fixatives; D007070:Immunoglobulin A; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D006735:Horseradish Peroxidase; D010447:Peptide Hydrolases; D005976:Glutaral