In vivo immunogold labeling confirms large-scale chromatin folding motifs.
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).
Kireev
Igor
I
Department of Cell and Developmental Biology, University of Illinois, Urbana-Champaign, 601 S. Goodwin Ave., Urbana, Illinois 61801, USA.
Lakonishok
Margot
M
Liu
Wenqiu
W
Joshi
Vishwas N
VN
Powell
Rick
R
Belmont
Andrew S
AS
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
D002843:Chromatin; D018809:Proliferating Cell Nuclear Antigen; D049452:Green Fluorescent Proteins