ARTICLE

An Evaluation of Antigen Retrieval Procedures for Immunoelectron Microscopic Classification of Amyloid Deposits

Correspondence to: Christoph Röcken, Inst. of Pathology, Otto-von-Guericke-University, Leipziger Str. 44, D-39120 Magdeburg, Germany.

The advantages of using immunoelectron microscopy in amyloid research and surgical pathology for the classification of amyloid deposits are well documented. The aim of this study was to improve single-labeling postembedding immunostaining by testing different antigen retrieval (AR) techniques. Etching and AR procedures were applied to sections from aldehyde-fixed and Epon-embedded autopsy specimens of patients who had suffered from generalized AA amyloidosis, systemic senile ATTR amyloidosis, or generalized -light chain amyloidosis. The procedures used were no AR, H₂O₂, saturated aqueous sodium metaperiodate (mPJ), heating in deionized water (dH₂O), heating in sodium citrate buffer (SCB), heating in EDTA (each 91C, 30 min), and combinations of etching and heating. Little effect was evident after treatment with H₂O₂, mPJ, and heating in dH₂O, but the signal density markedly increased after heating in 1 mM EDTA. Heating in SCB affected immunolabeling with anti-transthyretin and anti--light chain, whereas no effect was achieved for immunolabeling with anti-AA amyloid. We concluded that AR may significantly improve immunostaining of specimens that have undergone conventional fixation and embedding procedures for electron microscopy. The effect of AR on the detection of amyloid fibril proteins was probably mediated in part through chelation or binding of metal ions by the AR medium. (J Histochem Cytochem 47:1385–1394, 1999)

Key Words: amyloid, antigen retrieval, immunoelectron microscopy

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				J. Jendholm, M. Morgelin, M. L. A. Perez Vidakovics, M. Carlsson, H. Leffler, L.-O. Cardell, and K. Riesbeck Superantigen- and TLR-Dependent Activation of Tonsillar B Cells after Receptor-Mediated Endocytosis J. Immunol., April 15, 2009; 182(8): 4713 - 4720. [Abstract] [Full Text] [PDF]

				S. Yamashita, O. Katsumata, and Y. Okada Establishment of a standardized post-embedding method for immunoelectron microscopy by applying heat-induced antigen retrieval J. Electron Microsc. (Tokyo), March 30, 2009; (2009) dfp017v1. [Abstract] [Full Text] [PDF]

				C. Rocken, R. Kientsch-Engel, S. Mansfeld, B. Stix, K. Stubenrauch, B. Weigle, F. Buhling, M. Schwan, and W. Saeger Advanced Glycation End Products and Receptor for Advanced Glycation End Products in AA Amyloidosis Am. J. Pathol., April 1, 2003; 162(4): 1213 - 1220. [Abstract] [Full Text] [PDF]

				S. Yano, K. Kashima, T. Daa, S. Urabe, K. Tsuji, I. Nakayama, and S. Yokoyama An Antigen Retrieval Method Using an Alkaline Solution Allows Immunoelectron Microscopic Identification of Secretory Granules in Conventional Epoxy-embedded Tissue Sections J. Histochem. Cytochem., February 1, 2003; 51(2): 199 - 204. [Abstract] [Full Text] [PDF]

				B. Stix, T. Kahne, K. Sletten, J. Raynes, A. Roessner, and C. Rocken Proteolysis of AA Amyloid Fibril Proteins by Matrix Metalloproteinases-1, -2, and -3 Am. J. Pathol., August 1, 2001; 159(2): 561 - 570. [Abstract] [Full Text]

				S. Groos, E. Reale, and L. Luciano Re-evaluation of Epoxy Resin Sections for Light and Electron Microscopic Immunostaining J. Histochem. Cytochem., March 1, 2001; 49(3): 397 - 406. [Abstract] [Full Text]

				C. Rocken, B. Stix, D. Bromme, S. Ansorge, A. Roessner, and F. Buhling A Putative Role for Cathepsin K in Degradation of AA and AL Amyloidosis Am. J. Pathol., March 1, 2001; 158(3): 1029 - 1038. [Abstract] [Full Text] [PDF]

Guidelines | Subscriptions | About | exPRESS - Current - Archive | Business Information | Contact

The Journal of Histochemistry & Cytochemistry is owned, published, and licensed by The Histochemical Society © 1999