Figure 1: Workflow for visualizing the native ultrastructure of human motile cilia.
From: Cryo-electron tomography reveals ciliary defects underlying human RSPH1 primary ciliary dyskinesia
Human respiratory epithelial cells were obtained by nasal scrape biopsy, cultured, expanded and re-differentiated in an air–liquid interface culture. Ciliary axonemes were isolated using our method optimized for cryo-ET. Freshly isolated axonemes were rapidly frozen and transferred to a transmission electron microscope, where tilt series were recorded by stepwise rotation of the axonemes. Image processing was performed in silico to reconstruct the cryo-tomograms and increase the resolution by subtomogram averaging of the 96-nm repeats. The averaged native 3D structure of the human axonemal repeat can be visualized by isosurface rendering.
