4D Imaging of C. elegans Embryogenesis

Embryos were collected from gravid hermaphrodites and mounted with polystyrene beads (Polysciences Inc) as described (Du et al., 2015 (link)). Embryos were imaged on a Zeiss AxioObserver Z1 inverted microscope frame with Yokogawa CSU-X1 spinning disk and an Olympus UPLSAPO 60xs silicone oil immersion objective. GFP and mCherry channels were acquired simultaneously on a pair of aligned EMCCD cameras (C9100-13). Image acquisition was performed using MetaMorph software (Molecular Devices). Embryos were imaged every 75 s, with 30 z slices at 1 μm apart. Lineage tracing and quantification of marker expression were done with the StarryNite and AceTree software as described (Du et al., 2015 (link)).