title: In vivo immunogold labeling confirms large-scale chromatin folding motifs.
abstract: The difficulty in localizing specific cellular proteins by immuno-electron microscopy techniques limits applications of electron microscopy to cell biology. We found that in vivo immunogold labeling improves epitope accessibility, ultrastructural preservation and three-dimensional visualization, and allows correlated light and electron microscopy. We detected large-scale chromatin folding motifs within intact interphase nuclei of CHO cells and visualized the ultrastructure of DNA replication 'factories' labeled with GFP-proliferating cell nuclear antigen (PCNA).
authors.affiliation: Department of Cell and Developmental Biology, University of Illinois, Urbana-Champaign, 601 S. Goodwin Ave., Urbana, Illinois 61801, USA.
authors.forename: Igor
authors.initials: I
authors.lastname: Kireev
authors.forename: Margot
authors.initials: M
authors.lastname: Lakonishok
authors.forename: Wenqiu
authors.initials: W
authors.lastname: Liu
authors.forename: Vishwas N
authors.initials: VN
authors.lastname: Joshi
authors.forename: Rick
authors.initials: R
authors.lastname: Powell
authors.forename: Andrew S
authors.initials: AS
authors.lastname: Belmont
mesh_terms: D000818:Animals; D016466:CHO Cells; D002843:Chromatin / Q000648:ultrastructure*; D006224:Cricetinae; D003412:Cricetulus; D004261:DNA Replication / Q000502:physiology*; D049452:Green Fluorescent Proteins / Q000032:analysis; D007150:Immunohistochemistry; D007763:Lac Operon; D053768:Metal Nanoparticles; D008854:Microscopy, Electron / Q000379:methods*; D018809:Proliferating Cell Nuclear Antigen / Q000032:analysis
chemical_list: D002843:Chromatin; D018809:Proliferating Cell Nuclear Antigen; D049452:Green Fluorescent Proteins