Ti2e csu w1 spinning disc confocal system

The transgenic zebra sh strains scospondin-GFP ut24 and Tg (Foxj1:Arl13b-GFP) were created as previously described (Konjikusic, Yeetong et al. 2018 , Troutwine, Gontarz et al. 2020) . Tg(Foxj1:Arl13b-GFP); dnah10 ut28 and scospondin-GFP ut24 ;dnah10 ut28 lines were generated by cross homozygous dnah10 ut28 to each transgenic zebra sh and incrossing of their progeny to generate the homozygote. For live imaging, larvae were anaesthetized in 0.16% tricaine and embedded in 1% low-melt agarose. Time-lapse videos (from 10 s to 10 mins) were acquired at 0.1 to 33 Hz imaging frequency. Videos were cropped and edited by Adobe Premiere Pro (Version 12.0). Reissner ber and oor plate immuno uorescence staining was performed as previously described (Troutwine, Gontarz et al. 2020 ). In brief, samples were xed, permeabilized and incubated with primary antibodies. After washing and incubation with secondary antibodies, specimens were xed and lmed immediately. Confocal images and videos were taken using a Nikon Ti2E / CSU-W1 spinning disc confocal system. Images were analyzed and reconstructed using Fiji (Schindelin, Arganda-Carreras et al. 2012) . ROIs were manually adjusted on max intensity Z-projection.