title: Changes in chromatin structure during processing of wax-embedded tissue sections.
abstract: The use of immunofluorescence (IF) and fluorescence in situ hybridisation (FISH) underpins much of our understanding of how chromatin is organised in the nucleus. However, there has only recently been an appreciation that these types of study need to move away from cells grown in culture and towards an investigation of nuclear organisation in cells in situ in their normal tissue architecture. Such analyses, however, especially of archival clinical samples, often requires use of formalin-fixed paraffin wax-embedded tissue sections which need addition steps of processing prior to IF or FISH. Here we quantify the changes in nuclear and chromatin structure that may be caused by these additional processing steps. Treatments, especially the microwaving to reverse fixation, do significantly alter nuclear architecture and chromatin texture, and these must be considered when inferring the original organisation of the nucleus from data collected from wax-embedded tissue sections.
authors.affiliation: Breakthrough Breast Cancer Research Unit, Institute of Genetics and Molecular Medicine, University of Edinburgh, Crewe Road, Edinburgh, EH4 2XU, UK.
authors.forename: Elizabeth
authors.initials: E
authors.lastname: Kerr
authors.forename: Tomoharu
authors.initials: T
authors.lastname: Kiyuna
authors.forename: Shelagh
authors.initials: S
authors.lastname: Boyle
authors.forename: Akira
authors.initials: A
authors.lastname: Saito
authors.forename: Jeremy St J
authors.initials: JS
authors.lastname: Thomas
authors.forename: Wendy A
authors.initials: WA
authors.lastname: Bickmore
mesh_terms: D002467:Cell Nucleus / Q000235:genetics; D002478:Cells, Cultured / Q000378:metabolism*; D002843:Chromatin / Q000737:chemistry*; D006570:Heterochromatin / Q000502:physiology; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D016612:Paraffin Embedding; D016707:Tissue Fixation
chemical_list: D002843:Chromatin; D006570:Heterochromatin