Ou, Horng; Phan, Sébastien; Deerinck, Thomas J.; Thor, Andrea; Ellisman, Mark; O'Shea, Clodagh (2021). CIL:49951, Primary human small airway epithelial cells immortalized with telomerase. In Cell Image Library. UC San Diego Library Digital Collections. Dataset. https://doi.org/10.6075/J0862F1K

Multi-tilt EM tomogram of primary human small airway epithelial cell (SAEC) in interphase state. The tomogram refers to the SAEC in figure S7 of the published manuscript: http://science.sciencemag.org/content/357/6349/eaag0025http://science.sciencemag.org/content/357/6349/eaag0025.

• 2021

• Deerinck, Thomas J.
• Ellisman, Mark
• O'Shea, Clodagh
• Ou, Horng
• Phan, Sébastien
• Thor, Andrea

Primary human small airway epithelial cells in interphase state were fixed with 2.5% glutaraldehyde, 5mM 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 6 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 1.64 nm.

Preparation: Glutaraldehyde fixed cells, photo-oxidation of DRAQ5 in the presence of DAB solution to induce DAB polymerization and subsequent staining with osmium tetroxide
Relation to intact cell: semi-thin section ~250 nm
Item type: recorded image
Imaging mode: 8-tilt EM tomography
Parameter imaged: electron density
Source of contrast: Glutaraldehyde fixed cells, photo-oxidation of DRAQ5 in the presence of DAB solution to induce DAB polymerization and subsequent staining with osmium tetroxide

• Cell Image Library Group ID: 19851

• Primary human small airway epithelial cells immortalized with telomerase

• Primary human small airway epithelial cells in interphase state

• data
• image

• No linguistic content; Not applicable

Samplenumber: 49951

Doi: https://doi.org/10.6075/J0862F1K

Source data

• Source Record in the Cell Image Library: https://doi.org/10.7295/W9CIL49951

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2022-08-12