Near-isotropic, reconstructed volume electron microscopy (FIB-SEM) of human airway (cam_hum-airway-14771-b)

Sample: Human Primary Nasal Airway Epithelial Cells

Sample Description: This sample captures the cellular diversity of the human bronchial airway epithelium, the superficial layer of the respiratory tract whose major function is to provide a physical, biochemical and immunological barrier from the infection of viruses, bacteria and environmental pollutants inhaled through breathing.

Protocol: ALI filters with Karnovsky’s fixative in 0.1M sodium cacodylate buffer (SCB, pH 7.4) with 0.05% malachite green oxalate salt (M9015, Sigma-Aldrich Inc., MO USA). Post-fixed with the osmium-thiocarbohydrazide-osmium (OTO) method, with incubations in 1% osmium tetroxide in SCB for 1.5 hrs at RT, 2.5% potassium ferrocyanide in SCB for 1.5 hrs at RT, aqueous 1% thiocarbohydrazide solution for 45 mins at 40°C, and then with a 2% osmium tetroxide aqueous solution for 1.5 hrs 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 hrs at 50°C. After dehydration in ethanol/acetonitrile series, the sample was embedded in Epon/Araldite mixture.

Contributions: Samples were prepared by Nobu Morone (University of Cambridge) and acquired by Kirsty MacLellan-Gibson at the EM imaging facility at the Babraham Institute. ROI provided by Vito Mennella (University of Cambridge), sample provided by Chris O’Collaghan (UCL, GOSH), 3D segmentation by Aaran Vijayakumaran (University of Cambridge).

Acquisition ID: cam_hum-airway-14771-b

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

Data dimensions (µm): 44.00 x 20.04 x 21.42 (x, y, z)

Dataset URL: s3://janelia-cosem-datasets/cam_hum-airway-14771-b/cam_hum-airway-14771-b.zarr/recon-1/em/

Visualization Website: https://openorganelle.janelia.org/datasets/cam_hum-airway-14771-b

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