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Figure 2


Fig. 2. TEM appearance of Xi in WI-38 cells using conditions that preserve large-scale chromatin structure. (A) Deconvolved optical section from a live cell expressing histone H2B-GFP (top panel) shows a condensed Barr body (arrow) with a 200- to 400-nm-diameter substructure. Bottom panel: higher-magnification view of the Barr-body region, rotated 90° counterclockwise, from top panel view. (B-D) TEM images of 200-nm Epon sections through nuclei in samples prepared with three different fixation protocols all show a heterochromatic body with a similar 200- to 400-nm-wide chromatin substructure. (B) Direct fixation with 2% GA in phosphate buffer. (C) Cells in media pre-fixed by 8 minutes of UV irradiation in the presence of EtBr with a subsequent permeabilization step before fixation by 2% GA in buffer A*. (D) Fixation by 2% GA 1 minute after permeabilization in buffer A*. Higher-magnification views are shown at the bottom of each panel. Arrows indicate Xi heterochromatin body; Nu, nucleolus.





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