Research Project
Full description Dynamic remodeling of the actin cytoskeleton is a fundamental driver of epithelial morphogenesis during embryonic development. Here, we present a workflow for the acquisition and quantitative analysis of live confocal imaging data from transgenic Lifeact-EGFP quail embryos. The protocol combines long-term live imaging of the developing neural plate with automated image processing, apical surface extraction, Cellpose-based cell segmentation, cell tracking, and quantitative measurements of actin organization at single-cell resolution. The workflow enables identification of apically constricting cells based on temporal changes in apical area and quantification of F-actin dynamics within distinct cellular compartments, including the apical cortex and individual cell-cell junctions. In addition, junction length, orientation, and actin enrichment can be measured to distinguish different modes of actomyosin-mediated apical constriction. The protocol is compatible with widely used open-source software platforms and can be adapted to other epithelial tissues and fluorescent cytoskeletal reporters. By integrating live imaging with automated quantitative analysis, this workflow provides a robust framework for investigating the cellular and molecular mechanisms that drive neural tube morphogenesis and epithelial remodeling during development.