The data comprises bright-field time-lapse images of zebrafish embryos acquired in multiple batches within multi-well plates using an Acquifer Imaging Machine. Embryos from incrosses of Tg(sebox:EGFP) zebrafish strains were used for imaging. Imaging started with embryos between 2-3 hpf and ended with embryos between 27.25-28.25 hpf. Images of embryos were acquired at 248 acquisition time points with a time interval of 368 s between each acquisition time point. Individual embryo segments were identified and extracted using a trained neural network for object detection. Within this experiment folder, data are organized by microscope position and embryo number.