Z-series image of mouse ES cells carrying a beta-globin BAC transgene array (clone 8d) and transduced with a lentivirus containing an EGFP-lacI expression cassette. EGFP-LacI binding (green) to the 256mer lac operator repeat contained within the BAC reveals the location of the BAC transgene array relative to the nuclear periphery, highlighted by anti-nuclear pore immunostaining, and nuclear interior stained with DAPI (blue). A very high percentage of cells show part of the beta-globin transgene array touching the edge of the nuclear periphery.
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. Pixel size (xy) is 0.074 microns and z-spacing is 0.2 microns. 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.
|Spatial Axis||Image Size||Pixel Size|