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*CIL – Cell Image Library accession number. Please use this to reference an image.

CIL:26567*  Cite 
Description

Z-series images of mouse ES cells carrying DHFR BAC transgene array (clone 36) and differentiated into fibroblast-like cells. 3D FISH shows increase in number of lac operator foci (green) and change to a more chain / fiber like large-scale chromatin conformation for the BAC transgene array (red). DAPI staining is shown in blue. Lac operator sequences still segregate into foci but these are located within or peripheral to fiber-like segments visualized for the BAC FISH signal. Nonlinear folding of DNA sequences within linear large-scale chromatin "fiber" segments is implied by the appearance of these Lac operator repeat foci.

Technical Details

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 sizes are 0.074 microns (xy) 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.

Biological Sources
NCBI Organism Classification
Mus musculus
Cell Type
embryonic stem cell
Cellular Component
heterochromatin
nuclear chromatin
transgene
Biological Context
Biological Process
nucleus organization
chromatin organization
Attribution
Names
Paul Sinclair
Qian Bian
Matt Plutz
Edith Heard
Andrew Belmont
Published
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
Pubmed
20819934
Link
Journal of Cell Biology
Citation
Digital Object Identifier (DOI)
doi:10.7295/W9CIL26567
Archival Resource Key (ARK)
ark:/b7295/w9cil26567
Grouping This image is part of a group.
Imaging
Image Type
charge coupled device (CCD)
Image Mode
fluorescence microscopy
Parameters Imaged
fluorescence emission
Source of Contrast
distribution of a specific protein
differences in adsorption or binding of stain
Visualization Methods
4',6-diamidino-2-phenylindole (DAPI)
EGFP
Processing History
constrained iterative deconvolution
Data Qualifiers
processed data
suitable for spatial measurements
Sample Preparation
Methods
formaldehyde fixed tissue
Relation To Intact Cell
whole mounted tissue
Dimensions
Spatial Axis Image Size Pixel Size
X 512px 0.074µm
X 512px 0.074µm
Z 30px 0.2µm