This volume explores recent innovations across the entire pipeline of imaging signal transduction, cell preparation, cellular manipulation, image acquisition, and computational analysis. The chapters in this book cover topics such as rapid preparation of living Drosophila pupal macrophages for ex vivo imaging; controlling the potency of T cell activation using an optically tuneable chimeric antigen receptor; in situ imaging of proteins using DNA-PAINT super-resolution microscopy; reconstructing signaling networks using biosensor barcoding; and morphological, spatial, and dynamic models for cellular components. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Cutting-edge and authoritative, Imaging Cell Signaling is a valuable resource that provides guidance to researchers looking to learn how to effectively tailor design projects to image, manipulate, and model signal transduction.