A new immunoelectron microscopy approach for the detection of immunoglobulin and complement deposits in epoxy-embedded renal biopsies. 1998

S H Brorson, and E H Strøm
Department of Pathology, Ullevål Hospital, Oslo, Norway. s.h.brorson@ioks.uio.no

The purpose of this study was to examine the diagnostic value of a new immunoelectron microscopy technique (IEM) for detection of immunoglobulin and complement deposits in epoxy-embedded renal biopsies. Twenty-four renal biopsies were embedded in epoxy resin following a tissue processing involving moderately increased amount of accelerator, DMP-30 (Tri(Dimethyl Amino Methyl) Phenol), in the infiltration steps. Following antigen retrieval by heating in citrate buffer, immunogold labeling was performed on ultrathin sections from these epoxy blocks with antibodies against immunoglobulins and complement. The sections were counterstained with urnayl acetate and lead citrate without any enhancing procedures. The preservation of the ultrastructure with this method was similar to that usually seen in epoxy embedded material. The immunogold labeling was intense and distinct. Immunofluorescence (IF) for light microscopy was carried out on frozen sections of parallel tissue samples. The correspondence between IF and IEM were good, but in some cases higher sensitivity for IgA with IEM than IF was observed in the sense that smaller amounts of antigen were detectable with IEM. The combination of moderately increased amount of accelerator and antigen retrieval is superior to previous methods with respect to ease of use, ultrastructural preservation, and intensity of the immunolabeling. Moreover, the renal tissue can be processed in an automatic ultraprocessor together with other specimens which are to be prepared for routine electron microscopy.

UI MeSH Term Description Entries
D007070 Immunoglobulin A Represents 15-20% of the human serum immunoglobulins, mostly as the 4-chain polymer in humans or dimer in other mammals. Secretory IgA (IMMUNOGLOBULIN A, SECRETORY) is the main immunoglobulin in secretions. IgA,IgA Antibody,IgA1,IgA2,Antibody, IgA
D007678 Kidney Glomerulus A cluster of convoluted capillaries beginning at each nephric tubule in the kidney and held together by connective tissue. Glomerulus, Kidney
D004853 Epoxy Resins Polymeric resins derived from OXIRANES and characterized by strength and thermosetting properties. Epoxy resins are often used as dental materials. Epoxy Resin,Resin, Epoxy,Resins, Epoxy
D005921 Glomerulonephritis Inflammation of the renal glomeruli (KIDNEY GLOMERULUS) that can be classified by the type of glomerular injuries including antibody deposition, complement activation, cellular proliferation, and glomerulosclerosis. These structural and functional abnormalities usually lead to HEMATURIA; PROTEINURIA; HYPERTENSION; and RENAL INSUFFICIENCY. Bright Disease,Kidney Scarring,Glomerulonephritides,Scarring, Kidney
D006801 Humans Members of the species Homo sapiens. Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D000936 Antigen-Antibody Complex The complex formed by the binding of antigen and antibody molecules. The deposition of large antigen-antibody complexes leading to tissue damage causes IMMUNE COMPLEX DISEASES. Immune Complex,Antigen-Antibody Complexes,Immune Complexes,Antigen Antibody Complex,Antigen Antibody Complexes,Complex, Antigen-Antibody,Complex, Immune,Complexes, Antigen-Antibody,Complexes, Immune
D001707 Biopsy, Needle Removal and examination of tissue obtained through a transdermal needle inserted into the specific region, organ, or tissue being analyzed. Aspiration Biopsy,Puncture Biopsy,Aspiration Biopsies,Biopsies, Aspiration,Biopsies, Needle,Biopsies, Puncture,Biopsy, Aspiration,Biopsy, Puncture,Needle Biopsies,Needle Biopsy,Puncture Biopsies
D015932 Complement C3c A 206-amino-acid fragment in the alpha chain (672-1663) of C3b. It is generated when C3b is inactivated (iC3b) and its alpha chain is cleaved by COMPLEMENT FACTOR I into C3c (749-954), and C3dg (955-1303) in the presence COMPLEMENT FACTOR H. C3c Complement,Complement 3c,Complement C3c Fragment,Complement Component 3c,C3c Fragment, Complement,C3c, Complement,Complement, C3c,Component 3c, Complement,Fragment, Complement C3c
D016253 Microscopy, Immunoelectron Microscopy in which the samples are first stained immunocytochemically and then examined using an electron microscope. Immunoelectron microscopy is used extensively in diagnostic virology as part of very sensitive immunoassays. Immunoelectron Microscopy,Microscopy, Immuno-Electron,Immuno-Electron Microscopies,Immuno-Electron Microscopy,Immunoelectron Microscopies,Microscopies, Immuno-Electron,Microscopies, Immunoelectron,Microscopy, Immuno Electron
D019084 Fluorescent Antibody Technique, Indirect A form of fluorescent antibody technique commonly used to detect serum antibodies and immune complexes in tissues and microorganisms in specimens from patients with infectious diseases. The technique involves formation of an antigen-antibody complex which is labeled with fluorescein-conjugated anti-immunoglobulin antibody. (From Bennington, Saunders Dictionary & Encyclopedia of Laboratory Medicine and Technology, 1984) Immunofluorescence Antibody Test, Indirect,Immunofluorescence Technique, Indirect,Fluorescent Antibody Technic, Indirect,Immunofluorescence Technic, Indirect,Indirect Fluorescent Antibody Technic,Indirect Fluorescent Antibody Technique,Indirect Immunofluorescence,Indirect Immunofluorescence Assay,Assay, Indirect Immunofluorescence,Assays, Indirect Immunofluorescence,Immunofluorescence Assay, Indirect,Immunofluorescence Assays, Indirect,Immunofluorescence Technics, Indirect,Immunofluorescence Techniques, Indirect,Immunofluorescence, Indirect,Immunofluorescences, Indirect,Indirect Immunofluorescence Assays,Indirect Immunofluorescence Technic,Indirect Immunofluorescence Technics,Indirect Immunofluorescence Technique,Indirect Immunofluorescence Techniques,Indirect Immunofluorescences

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