Publication: Antigen retrieval on epoxy sections based on tissue infiltration with a moderately increased amount of accelerator to detect immune complex deposits in glomerular tissue
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Date
1999
Authors
Brorson, S.H. ; Andersen, T. ; Haug, S. ; Kristiansen, I. ; Risstubben, A. ; Tchou, H. ; Ulstein, J.
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Publisher
Murcia : F. Hernández
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DOI
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info:eu-repo/semantics/article
Description
Abstract
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(Dimethy1 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. Immunogold labeling was performed on
epoxy sections and LR-White sections of renal tissue
with IgG-deposits, and the antibody used was anti-IgG.
Antigen retrieval was performed by heating the sections
in citrate buffer. The amount of immunogold labeling on
retrieved sections increased according to the amount of
accelerator the non-osmicated epoxy sections were based
on in the infiltration steps. For the osmicated epoxy
sections these differences were less pronounced. The
immunogold labeling of retrieved epoxy sections was up
to 70% of LR-White labeling. In addition to breaking
fixation bond introduced by the chemical fixation, we
believe that the antigen retrieval also breaks bonds
between the epoxy resin and the embedded tissue. The
combination of increased amount of accelerator in the
tissue infiltration and antigen retrieval by heating the
sections in citrate buffer is a good method for improving
the immunolabeling of epoxy sections.
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