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

CIL:49803*  Cite 

Multi-tilt EM tomogram of a human bone osteosarcoma (U2OS) cell in the mitotic anaphase state. The tomogram refers to the U2OS cell in Figure 7 of the published manuscript:

Technical Details

A human bone osteosarcoma cell in anaphase mitotic state was fixed with 2.5% glutaraldehyde, 5 mM CaCl2 in 0.1 M sodium cacodylate buffer (pH 7.4). Cells were stained with 10 µM DRAQ5, and photo-oxidized at 620 ± 30 nm for 3 minutes. Cells were post-fixed with 2% osmium tetroxide, 2 mM CaCl2, 1.5% potassium ferrocyanide in 0.15 M sodium cacodylate buffer (pH 7.4). Samples were ethanol dehydrated, embedded in Durcupan epoxy resin and sectioned to 250 nm thickness and mounted on copper slot grid. 5 nm and 10 nm colloidal gold particles were deposited on the grid as fiducial markers. 8-tilt EM tomography was collected on a 300 kV FEI Titan Krios using a 4K by 4K Gatan Ultrascan CCD camera at 1 degree increment for each tilt series from -60° to +60°. The entire set of EMT images was aligned and reconstructed using the Transform-based Tracking, Bundle adjustment, and Reconstruction (TXBR) package. The data has a voxel size of 1.28 nm.

Biological Sources
NCBI Organism Classification
Human bone osteosarcoma cells
Biological Context
Biological Process
A human bone osteosarcoma cell in anaphase mitotic state
Horng Ou
Sebastien Phan
Thomas Deerinck
Andrea Thor
Mark Ellisman
Clodagh O’Shea
Digital Object Identifier (DOI)
Archival Resource Key (ARK)
Grouping This image is part of a group.
Image Type
recorded image
Image Mode
8-tilt EM tomography
Parameters Imaged
electron density
Source of Contrast
Osmium tetroxide staining of DAB polymers catalyzed by photo-oxidation of DRAQ5 stained cells in the presence of DAB solution
Sample Preparation
Glutaraldehyde fixed cells, photo-oxidation of DRAQ5 in the presence of DAB solution to induce DAB polymerization and subsequent staining with osmium tetroxide
Spatial Axis Image Size Pixel Size
X 800px 1.28nm
Y 1000px 1.28 nm
Z 1000px 1.28 nm