Re-evaluation of epoxy resin sections for light and electron microscopic immunostaining.
Epoxy resins provide optimal tissue morphology at both the light and the electron microscopic level and therefore enable correlative studies on semithin and thin sections from the same tissue block. Here we report on an approach to retain these advantages for immunolabeling studies by adapting and combining well-known techniques, i.e., surface etching with sodium ethoxide and heat-mediated antigen retrieval. We propose a simple procedure for immunostaining semithin and thin epoxy resin sections. To check its applicability, well characterized, commercially available antibodies (against E-cadherin, alpha-catenin, and beta-catenin) were used on sections of human small intestine. By light microscopy, the immunostaining efficiency was compared on cryo-, paraffin, and epoxy semithin sections processed in parallel. The most detailed results were obtained on semithin sections, where the labeling precisely delineated the lateral plasma membrane of the enterocytes. At the electron microscopic level the procedure did not damage the structures and allowed an efficient, reproducible immunogold labeling extending homogeneously over exceptionally wide tissue areas. The three antibodies specifically labeled the zonula adherens of the junctional complex between epithelial cells and, in agreement with light microscopic observations, the lateral plasma membrane.
Groos
S
S
Center of Anatomy, Hannover Medical School, Hannover, Germany. groos.stephanie@mh-hannover.de
Reale
E
E
Luciano
L
L
D015820:Cadherins / Q000378:metabolism; D003598:Cytoskeletal Proteins / Q000378:metabolism; D004853:Epoxy Resins; D006801:Humans; D007150:Immunohistochemistry; D007583:Jejunum / Q000378:metabolism; D008027:Light; D008853:Microscopy / Q000379:methods*; D016253:Microscopy, Immunoelectron / Q000379:methods; D016613:Plastic Embedding; D015534:Trans-Activators; D051178:alpha Catenin; D051176:beta Catenin
C495273:CTNNA1 protein, human; C495270:CTNNB1 protein, human; D015820:Cadherins; D003598:Cytoskeletal Proteins; D004853:Epoxy Resins; D015534:Trans-Activators; D051178:alpha Catenin; D051176:beta Catenin