diSPIM imaging

Imaging was conducted with a dual-view inverted selective plane illumination microscope (diSPIM) as described (Kumar et al., 2014). The microscope was equipped with LMM5 laser (Laser Illumination Laser Merge Module 5, Spectral Applied Research, Richmond Hill, Canada) and Quad Filterset (F59-405/F73-410/F57-406) purchased from AHF (Tübingen, Germany). Images were acquired by two water-cooled ORCA-Flash4.0 Hamamatsu sCMOS cameras (Hamamatsu, Japan). Cooling was provided with Julabo F250 cooling circuit (Julabo, Seelbach, Germany). Standard culture conditions were provided by an incubation chamber (3i ECS2) from Intelligent Imaging Innovations (Denver, CO); direct airflow over the SPIM head was minimized to avoid unnecessary vibrations. All imaging time lapse acquisitions were conducted with 320 µW laser power for 488 nm excitation wavelength (measured at the sample). Readjusting the fine alignment of the microscope was conducted shortly before start of the acquisition.