Cell polarity signaling : methods and protocols Use of Fluorescence Recovery after Photobleaching (FRAP) to Measure In Vivo Dynamics of Cell Junction Associated Polarity Proteins FRET Imaging of Rho GTPase Activity with Red Fluorescent Protein Based FRET Pairs Live‐Cell Total Internal Reflection Fluorescence (TIRF) Microscopy to Investigate Protein Internalization Dynamics Single Cell Single Molecule Pull Down (sc SiMPull) for Detection of Protein Complexes from Embryonic Lysates Biochemical Assays to Detect Activation of Small GTPases Rho Rac and Cdc42 during Morphogenesis Analysis of Planar Cell Polarity Complexes by Proximity Biotinylation in Xenopus Embryos Regulation of Cell Polarity by Posttranslational Protein Palmitoylation Enrichment and Detection of Wnt Proteins from Cell Culture Media Using Live Imaging to Examine Early Cardiac Development in Zebrafish Imaging Planar Cell Polarity Proteins in Xenopus Neuroectoderm In Ovo Gain and Loss of Function Approaches to Study Gut Morphogenesis Avian Embryos as a Model to Study Vascular Development BAC Recombineering and Transgenesis to Study Cell Polarity and Polarized Tissue Morphogenesis in Mice Two Photon Cell and Tissue Level Laser Ablation Methods to Study Morphogenetic Biomechanics Protocols for Investigating the Epithelial Properties of Cardiac Progenitor Cells in the Mouse Embryo Methods to Investigate Cell Polarity of Inner Ear Characterization of Axon Guidance Phenotypes in Wnt PCP Mutant Mice In Vitro Explant Assays and Cultures to Study PCP Signaling in Axon Guidance Biochemical and Cellular Assays to Study Mechanisms of PCP Signaling in Axon Guidance Dissection Fixation and Immunostaining of the Drosophila Midgut In Vivo Analysis of Pathways Regulating Epithelial Polarity and Secretion Using Drosophila Salivary Glands Imaging Epidermal Cell Rearrangement in the C. elegans Embryo Methods for the Study of Apical Constriction during Ascidian Gastrulation Assays for Apical Constriction Using the Xenopus Model The Use of 3 Dimensional Cell Culture to Study Apicobasal Polarization and Lumen Formation Studying Cell Polarity Dynamics during Cancer Initiation Using Inducible 3D Organotypic Cultures Under Agarose Chemotaxis and Migration Assays for Dictyostelium Mapping Asymmetry in Collective Cell Migration: Lessons from Border Cells in Drosophila Oogenesis Targeted Injections Atomic Force Microscopy and Hydrogels: A Toolbox to Study Tissue Mechanics In Vivo and Ex Vivo Quantitative Analysis of Directional Neural Crest Cell Migration. This volume explores detailed methods to investigate various aspects of biology related to cell polarity or asymmetry within a cell. Molecular cellular and tissue level regulation and function as well as diseases caused by impairment of cell polarity are explored by these methods. Beginning with advanced imaging and biochemical methods the book continues with planar cell polarity (PCP) signaling in morphogenesis in diverse developmental contexts apicobasal (AB) cell polarity in development and diseases as well as directional cell migration and biomechanics in cell polarity. The collection includes the usage of a wide variety of model systems and an extensive array of techniques including genetic imaging biochemical and biomechanical. Written for the highly successful Methods in Molecular Biology 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. Authoritative and comprehensive Cell Polarity Signaling: Methods and Protocols aims to enable researchers to delve into the stimulating field of cell polarity and contribute to our understanding of how coordinated control of protein stability trafficking membrane retention post translational modification and dynamic organization leads to active regulation of cell polarity. Chapter 29 is available open access under a Creative Commons Attribution 4.0 International License via link.springer.com.