A method for measurements of the efficiency of immunogold labelling of epoxy-embedded proteins subjected to different retrieval techniques.

The purpose of this study was to compare the level of immunogold labelling of both osmicated and non-osmicated epoxy sections when subjected to different antigen retrieval, etching and incubation temperature for the antibodies. Pure IgG protein gels were produced by glutaraldehyde fixation, eventually postfixed with 1% osmium tetroxide, and embedded in epoxy resin. Ultrathin sections were antigen retrieved in citrate solution at 95 or 144 degrees C and eventually etched with NaIO4.

Increased level of immunogold labeling of epoxy sections by rising the temperature significantly beyond 100 degrees C in the antigen retrieval medium.

The purpose of this study was to compare the level of immunogold labeling of epoxy sections when the sections were subjected to antigen retrieval at different temperatures. Renal swine tissue with glomerular immune complex deposits with reactivity against IgG and C3 was embedded in epoxy resin. Sections from these blocks were exposed to antigen retrieval by heating in citrate solution at temperatures in the range of 25-135 degrees C.

A comparative study of the immunogold labeling on H(2)O(2)-treated and heated epoxy sections.

The purpose of this study was to compare the intensity of the immunogold labeling of H(2)O(2)-treated and heated epoxy sections. Renal swine tissue with glomerular immune complex deposits with reactivity against IgG was embedded in epoxy resin. Immunogold labeling with anti-IgG was performed on sections from these blocks.

Deplasticizing or etching of epoxy sections with different concentrations of sodium ethoxide to enhance the immunogold labeling.

The study's purpose was to obtain improved "deplasticizing" of epoxy sections for immunoelectron microscopy. Epoxy-embedded renal swine tissue with immune complex deposits was used. Ultrathin sections were mounted on uncoated grids or on carbon-stabilized formvar grids.

Fixative-dependent increase in immunogold labeling following antigen retrieval on acrylic and epoxy sections.

We examined the increase in immunogold labeling of variably fixed, resin embedded tissue sections following antigen retrieval by heating in citrate solution. Fibrin clots and porcine renal tissue were fixed in glutaraldehyde, paraformaldehyde or ethanol, and specimens were embedded in LR-White or epoxy resin. Immunogold labeling was performed on ultra-thin sections with anti-fibrinogen for the fibrin clots and anti-IgG for the porcine renal tissue.

Increased yield of immunogold labeling of epoxy sections by adding para-phenylendiamine in the tissue processing.

The purpose of this study was to examine if the presence of para-phenylendiamine (PPD) in the tissue processing could increase the yield of immunogold labeling of the epoxy sections. Renal swine tissue with glomerular immune complex deposits with reactivity against IgG was embedded in epoxy resin. PPD was added (1) at the beginning of the dehydration, (2) in the first step with propylene oxide, (3) in the beginning of the dehydration and in all steps with propylene oxide included the infiltration step where propylene oxide and epoxy resin are mixed, or (4) PPD was totally avoided.

Comparison of the immunolabeling of heated and deplasticized epoxy sections on the electron microscopical level.

The purpose of this study was to compare the yield of immunogold labeling of heated epoxy sections with the yield of labeling of deplasticized epoxy sections, and to compare the immunolabeling of deplasticized high-accelerator epoxy sections and deplasticized low-accelerator epoxy sections. Renal swine tissue and human thyroid tissue were embedded in both high- and low-accelerator epoxy resin and also in LR-White resin. Immunogold labeling was performed on deplasticized (ethoxide-treated), heated and non-treated ultrathin sections from these specimens.

Comparison of the immunogold labeling of single light chains and whole immunoglobulins with anti-kappa on LR-white and epoxy sections.

The purpose of this study was to examine the intensity of the immunogold labeling of kappa light chains as single molecules and as parts of whole immunoglobulin molecules in LR-White sections and epoxy sections both practically and theoretically. Human renal tissues including deposits of kappa light chains and immune complex deposits of IgA were embedded in both LR-White resin and epoxy resin. Immunogold labeling was performed on unetched thin sections of both resins with anti-kappa or anti-IgA.

Antigen retrieval on epoxy sections based on tissue infiltration with a moderately increased amount of accelerator to detect immune complex deposits in glomerular tissue.

We wanted to examine the effect of antigen retrieval on epoxy sections where the tissue had been infiltrated by resin containing moderately increased amounts of accelerator. The concentration of accelerator DMP-30 (Tri(Dimethyl Amino Methyl) Phenol) was varied in the range of 0% to 4% in the infiltration step of the tissue processing. Some of the epoxy sections were fixed in osmium tetroxide, and for others this fixative was avoided.

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

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.

Bovine serum albumin (BSA) as a reagent against non-specific immunogold labeling on LR-White and epoxy resin.

The purpose of this study was to examine how different incubation times with different concentrations of bovine serum albumin (BSA) affect the amount of non-specific immunogold labeling on epoxy sections and LR-White sections. Immunogold labeling was performed on epoxy sections and LR-White sections of renal tissue with IgG-deposits and fibrin clots, and the antibodies used were anti-IgG and anti-fibrinogen, respectively. The sections were incubated with different concentrations of BSA prior to application of primary antibodies, and the length of this pre-incubation step varied between 0 and 4 h.