Nature Methods
- 5, 311 - 313 (2008)
Published online: 16 March 2008; | doi:10.1038/nmeth.1196
In vivo immunogold labeling confirms large-scale chromatin folding motifsIgor Kireev1, 2, Margot Lakonishok1, 4, Wenqiu Liu3, Vishwas N Joshi3, Rick Powell3 & Andrew S Belmont11
Department of Cell and Developmental Biology, University of Illinois, Urbana-Champaign, 601 S. Goodwin Ave., Urbana, Illinois 61801, USA. 2
Department of Electron Microscopy, A.N. Belozersky Institute of Physico-Chemical Biology, Moscow State University, Building A, Leninskie Gory, GSP-1, Moscow 119991, Russia. 3
Nanoprobes, Inc., 95 Horse Block Road, Unit 1, Yaphank, New York 11980, USA. 4
Present address: Department of Molecular and Cell Biology, Northwestern Medical School, 303 E. Chicago Ave., Ward 11-080, Chicago, Illinois 60611, USA.
Correspondence should be addressed to Andrew S Belmont asbel@uiuc.edu 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).
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