Video Dataset of Mouse Trachea Respiratory Cilia (Control and following Hydrogen Peroxide treatment)

Ciliated tracheal epithelial samples were collected for video microscopy as described previously (PMID: 41149796). Samples were incubated for 10 min in L-15 media + 10% FBS containing different Hydrogen Peroxide concentrations (0.1%, 0.2%, 0.5%, 1%). L-15 media + 10% FBS without Hydrogen Peroxide served as the control. For imaging, tracheal samples were mounted with ~100 µL L15+10%FBS in a shallow imaging chamber (PMID: 41149796). 1µm microspheres (Polysciences, Warrington, PA; 08226-15) were added to the bathing media to track cilia-driven flow. Tracheal samples were imaged at 37.5 °C on a Zeiss Axiovert 200 microscope using DIC and 63×/1.4 Oil objective. Ciliary motion was captured using a high speed USB3.0 digital camera with a 1.5MP 1/2.9” Sony Exmore CMOS sensor (ProSciTech; EM101500A). One second movies @>250fps (uploaded here to zenodo) were collected to determine cilia beat frequency (CBF) after 10 minutes treatment. Image pixel size = 0.104 micrometers.