Near-isotropic, reconstructed volume electron microscopy (FIB-SEM) of human airway (cam_hum-airway-14500)

Sample: Human Primary Nasal Airway Epithelial Cells

Sample Description: Human Airway Epithelium, from immortalised basal stem cell progenitors, differentiated using Air Liquid Interphase (ALI) cultures

Protocol: Differentiated human airway cells cultures grown on ALI filters were fixed with Karnovsky’s fixative in 0.1 M sodium cacodylate buffer (SCB, pH 7.4) with 0.05 % malachite green oxalate salt (M9015, Sigma-Aldrich Inc., MO USA). Briefly, specimens were post-fixed with the osmium-thiocarbohydrazide-osmium (OTO) method, with incubations in 1% osmium tetroxide in SCB for 1.5 hr at RT, 2.5% potassium ferrocyanide in SCB for 1.5 hr at RT, aqueous 1% thiocarbohydrazide solution for 45 min at 40 °C, and then with a 2% osmium tetroxide aqueous solution for 1.5 hr at RT, followed by en-bloc staining with aqueous 1% uranyl acetate solution overnight at 4 °C. Samples were subsequently transferred in lead aspartate solution (pH 5.0) for 2 hr at 50 °C. After dehydration in ethanol/acetonitrile series, the sample was embedded in Epon/Araldite mixture.

Contributions: Samples were prepared and acquired at the University of Montreal EM imaging facility, region of Interest (ROI) sample provided by Vito Mennella (University of Cambridge), 3D segmentation by Aaran Vijayakumaran (University of Cambridge).

Acquisition ID: cam_hum-airway-14500

Voxel size (nm): 10 x 10 x 10 (x, y, z)

Data dimensions (µm): 15.36 x 10.24 x 74.30 (x, y, z)

Dataset URL: s3://janelia-cosem-datasets/cam_hum-airway-14500/cam_hum-airway-14500.zarr/recon-1/em/

Visualization Website: https://openorganelle.janelia.org/datasets/cam_hum-airway-14500

Publication: Xu et al., 2017; Vijayakumaran, et al., 2024