In vivo immunogold labeling confirms large-scale chromatin folding motifs.

Kireev, Igor; Lakonishok, Margot; Liu, Wenqiu; Joshi, Vishwas N; Powell, Rick; Belmont, Andrew S · Nat Methods · 2008

basic_science · Level V

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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).

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