title: Isolation of Dictyostelium nuclei for light and electron microscopy.
abstract: The significance of amoebae for studies of nuclear architecture has considerably increased in the recent years. The availability of a protocol for isolation of nuclei in a quality sufficient for high-resolution light and electron microscopy is a prerequisite for such studies. Here we present a protocol for high enrichment of nuclei by sucrose density-gradient centrifugation. Moreover, we describe how to use these isolated nuclei as specimens for immunofluorescence and immune-electron microscopy of ultrathin sections.
authors.affiliation: Department of Cell Biology, Institute for Biochemistry and Biology, University of Potsdam, Potsdam-Golm, Germany.
authors.forename: Petros
authors.initials: P
authors.lastname: Batsios
authors.forename: Otto
authors.initials: O
authors.lastname: Baumann
authors.forename: Ralph
authors.initials: R
authors.lastname: Gräf
authors.forename: Irene
authors.initials: I
authors.lastname: Meyer
mesh_terms: D002458:Cell Fractionation / Q000379:methods*; D002467:Cell Nucleus / Q000378:metabolism / Q000648:ultrastructure*; D002499:Centrifugation, Density Gradient / Q000379:methods; D004023:Dictyostelium / Q000166:cytology* / Q000378:metabolism / Q000648:ultrastructure; D005404:Fixatives / Q000737:chemistry; D019084:Fluorescent Antibody Technique, Indirect; D016253:Microscopy, Immunoelectron / Q000379:methods; D009685:Nuclear Envelope / Q000378:metabolism / Q000648:ultrastructure; D016613:Plastic Embedding
chemical_list: D005404:Fixatives