Light-sheet fluorescence imaging charts the gastrula origin of vascular endothelial cells in early zebrafish embryos

摘要

Parental crossing: collection of transgenic embryos from crosses between Tg(H2A.F/Z:EGFP) and Tg(kdrl:mCherry) transgenic lines. Sample mounting: careful de-chorionation of embryos at 4 hpf and their transfer into a fluorinated ethylene propylene tube filled with 0.2% agarose. The tube is fixed to a fine wire and then mounted to a holder. Sample imaging: The illumination objectives (IOs) illuminate the sample from the left and right alternately, while the detection objective (DB) detects signals at 0°. The embryo is then rotated and the 180° images are acquired in the same way. Image processing: The raw data (~7 TB) are processed using the AFEIO software to obtain fused high-resolution data (~70 GB). Data analysis: The processed data are imported into Imaris software to run retrospective lineage analysis and determine a gastrula map of the origin of vascular ECs from 6 to 27 hpf.