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World J Gastroenterol. Jun 21, 2010; 16(23): 2851-2866
Published online Jun 21, 2010. doi: 10.3748/wjg.v16.i23.2851
Fixation methods for electron microscopy of human and other liver
Eddie Wisse, Filip Braet, Hans Duimel, Celien Vreuls, Ger Koek, Steven WM Olde Damink, Maartje AJ van den Broek, Bart De Geest, Cees HC Dejong, Chise Tateno, Peter Frederik
Eddie Wisse, Hans Duimel, Celien Vreuls, Peter Frederik, Electron Microscope Unit, University of Maastricht, 6200 MD Maastricht, The Netherlands
Eddie Wisse, Filip Braet, Australian Key Center for Microscopy and Microanalysis, Sydney University, NSW 2006 Sydney, Australia
Eddie Wisse, Ger Koek, Department of Internal Medicine, University of Maastricht, 6200 MD Maastricht, The Netherlands
Steven WM Olde Damink, Maartje AJ van den Broek, Cees HC Dejong, Department of Surgery, University of Maastricht, 6200 MD Maastricht, The Netherlands
Bart De Geest, Center for Molecular and Vascular Biology, University of Leuven, 3000 Leuven, Belgium
Chise Tateno, PhoenixBio Co., 3-4-1 Kagamiyama, Higashi-Hiroshima 739-0046, Japan
Author contributions: Wisse E and Braet F designed the fixation procedures and wrote the manuscript together; Wisse E performed the EM observations; Braet F provided the schematic diagrams; Duimel H and Vreuls C prepared the specimens and sections and provided technical support; Olde Damink SWM, Dejong CHC, van den Broek MAJ and Koek G provided the human liver wedge biopsies, obtained approval of the ethical committee and provided the clinical data; De Geest B provided rabbit, mouse and human livers and biopsies; Tateno C provided mouse livers of different strains; Frederik P supervised the study as head of the EM Unit.
Correspondence to: Dr. Eddie Wisse, Professor, Electron Microscope Unit, University of Maastricht, PO Box 616, 6200 MD Maastricht, The Netherlands. eddie@wisse.be
Telephone: +31-43-3881280 Fax: +31-43-3671040
Received: December 29, 2009
Revised: January 25, 2010
Accepted: February 1, 2010
Published online: June 21, 2010
Abstract

For an electron microscopic study of the liver, expertise and complicated, time-consuming processing of hepatic tissues and cells is needed. The interpretation of electron microscopy (EM) images requires knowledge of the liver fine structure and experience with the numerous artifacts in fixation, embedding, sectioning, contrast staining and microscopic imaging. Hence, the aim of this paper is to present a detailed summary of different methods for the preparation of hepatic cells and tissue, for the purpose of preserving long-standing expertise and to encourage new investigators and clinicians to include EM studies of liver cells and tissue in their projects.

Keywords: Liver; Tissue fixation; Perfusion; Electron microscopy; Biopsy