Visualizing mouse embryo gastrulation epithelial-mesenchymal transition through single cell labeling followed by ex vivo whole embryo live imaging

Abstract

Epithelial-mesenchymal transition (EMT) is often studied in pathological contexts, such as cancer or fibrosis. This chapter focuses on physiological EMT that allows the separation of germ layers during mouse embryo gastrulation. In order to record individual cells behavior with high spatial and temporal resolution live imaging as they undergo EMT, it is very helpful to label the cells of interest in a mosaic fashion so as to facilitate cell segmentation and quantitative image analysis. This protocol describes the isolation, culture, and live imaging of E6.5–E7.5 mouse embryos mosaically labeled in the epiblast, the epithelium from which mesoderm and endoderm layers arise through EMT at gastrulation.