Cell-cell communication plays a pivotal role in coordination and function of biological systems. In this context, we want to investigate the relationships between the dynamics of kinase activities and molecular mechanics in living tissues with high sensitivity and spatiotemporal resolution. To this aim, we will use an approach relying on multiplexed functional live cell imaging with various genetically-encoded Kinase Translocation Reporters and custom genetically-encoded FRET biosensors. We will first use this approach in 2D tissues and then in 3D cellular models, which provide venues to investigate morphogenesis and for drug discovery, as their architecture mimics native in vivo microenvironments. At the end of this workshop, participants will be able to generate rich multiparametric images of basic biological processes. We will also perform on-the-fly quantitative image analysis of generated datasets by using custom-made routines developed on ImageJ to automatically segment cells and track their biosensors outputs over time.