Near-isotropic, reconstructed volume electron microscopy (FIB-SEM) of human airway (jrc_hum-airway-14953vc)
Sample: Human Airway Epithelium, from immortalised basal stem cell progenitors, differentiated using Air Liquid Interphase (ALI) cultures
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: Human multiciliated cells cultures differentiated for 28 days were fixed with Karnovsky’s fixative in 0.1M sodium cacodylate buffer (SCB, pH 7.4) with 0.05% malachite green oxalate salt and processed largely as previously described (Liu et al., 2020; Morone et al., 2024). Briefly, specimens were 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 TAAB E202(/1) Araldite/TAAB 812 Epon mixture Resin.
Contributions: Region of Interest (ROI) and biology description provided by Vito Mennella (University of Cambridge), sample provided by Vito Mennella, Nobu Morone and Sophie Breugesem (University of Cambridge), prepared for imaging by Zhiyuan Lu (HHMI/Janelia), with imaging by Wei Qiu (HHMI/Janelia) and Christopher Bleck (HHMI/Janelia), post-processing by Eric Trautman, Michael Innerberger and Stephan Preibisch (HHMI/Janelia).
Acquisition ID: jrc_hum-airway-14953vc
Voxel size (nm): 8 x 8 x 8 (x, y, z)
Data dimensions (µm): 212.94 x 103.02 x 120.74 (x, y, z)
Imaging start date: 2023-11-21
Imaging duration (days): 32
Dataset URL: https://data.janelia.org/gV96S
Visualization Website: https://openorganelle.janelia.org/datasets/jrc_hum-airway-14953vc
Publication: Xu et al., 2017; Vijayakumaran, et al., 2024
