z-series image of mouse ES cells with a multi-copy alpha-globin BAC insertion. H3K9me3 (blue) marks the heterochromatic core which largely colocalizes with combined FISH signal from the lac operator repeats and BAC vector backbone (green). In contrast the BAC FISH signal (red) is more peripheral.
Formaldehyde fixed cells were imaged using an IMT-2 Olympus fluorescence microscope with 60x 1.4 NA objective lens. Z-series were recorded, and the slices processed by iterative constrained deconvolution. xy pixel dimensions are 0.067 microns, z spacing is 0.2 microns and number of z-slices 14. See also: P Sinclair et al. 2010 Dynamic plasticity of large scale chromatin structure revealed by self-assembly of engineered chromosome regions. J Cell Biol 109:761-776.
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