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. Some of these sections were treated by H(2)O(2), others were heated in a citrate solution, while some were not treated at all. Some epoxy sections, which had been exposed to both H(2)O(2) and heat, were also exposed to the same immunolabeling. The heated epoxy sections obtained an yield of specific immunogold labeling, which was twice as large as the labeling of the H(2)O(2)-treated sections. The yield of immunolabeling of the sections that had been exposed to both H(2)O(2) and heat was not significantly different from the sections that were only exposed to heat. The non-treated sections were very weakly labeled with anti-IgG. We believe that both H(2)O(2) and heat have the ability to break some chemical bonds between the epoxy resin and the antigens, but heating in citrate buffer has a larger potential in this respect than H(2)O(2). We interpret the results from the combined treatment with H(2)O(2) and heat in the following way; the bonds that are broken by H(2)O(2) will also be broken by heating in citrate solution. The practical significance of these results is that heating in citrate buffer is a more convenient method for enhancing the immunolabeling of epoxy sections than treatment with H(2)O(2).
Department of Pathology, Ullevål Hospital, Kirkeveien 166, 0407 Oslo, Norway. s.h.brorson@ioks.uio.no
S H
SH
Brorson
A R
AR
Hansen
H Z
HZ
Nielsen
I K
IK
Woxen
D000818:Animals; D000936:Antigen-Antibody Complex / Q000302:isolation & purification*; D004853:Epoxy Resins; D006046:Gold; D016591:Histocytological Preparation Techniques; D006861:Hydrogen Peroxide; D007672:Kidney Cortex / Q000648:ultrastructure*; D007678:Kidney Glomerulus / Q000648:ultrastructure*; D016253:Microscopy, Immunoelectron / Q000379:methods*; D013552:Swine
D000936:Antigen-Antibody Complex; D004853:Epoxy Resins; D006046:Gold; D006861:Hydrogen Peroxide