Mechanobiology Methods and Protocols

This detailed book collects methodologies exploring mechanobiology, the involvement of mechanical forces in cell fate specification and in controlling single and collective cell behaviors such as directed migration, morphogenesis, wound healing, and the immune response. The volume features methods to quantify the mechanical properties of cells and adhesion proteins, to expose cells to external mechanical forces, to quantitatively characterize mechano-responses at various scales, to measure forces applied by cells on the extracellular matrix, as well as chapters on force measurement inside cells, probing cell signaling and gene expression in response to force, and biophysical modeling of cell shape and protein dynamics. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary material and reagents, step-by-step and readily reproducible protocols, and tips on troubleshooting and avoiding known pitfalls. 

Measuring Cell Mechanical Properties Using Microindentation.- In Situ Measurements of Cell Mechanical Properties Using Force-Spectroscopy.- Quantification of Apparent Membrane Tension and Membrane-to-Cortex Attachment in Animal Cells Using Atomic Force Microscopy-Based Force Spectroscopy.- Characterizing the Biophysical Properties of Adhesive Proteins in Live Cells Using Single Molecule Atomic Force Microscopy.- Application of Shear Stress to Endothelial Cells Using a Parallel Plate Flow Chamber.- Cell Stretcher Assay to Analyze Mechanoresponses to Cyclic Stretching.- Two-Point Optical Manipulation of Cell Junctions in the Early Epithelium of the Drosophila Embryo.- A Microfluidic-Like System (MLS) to Grow, Image, and Quantitatively Characterize Rigidity-Sensing by Plant’s Roots and Root Hair Cells.- Photoresponsive Hydrogels for Studying Mechanotransduction of Cells.- Analyzing Mechanically Evoked Currents at Cell-Substrate Junctions.- Quantifying Strain Sensing Protein Recruitment During Stress Fiber Repair.- Quantification of Invadopodia Formation and Matrix Degradation Activity.- Measuring Cellular Traction Forces with Micropillar Arrays.- Imaging Cell Adhesive Force at the Single Molecule Level.- Multiplexed Molecular Tension Sensor Measurements Using PIE-FLIM.- Visualizing Neurons under Tension In Vivo with Optogenetic Molecular Force Sensors.- Single-Cell Quantification of the Mechanical Stability of Cell-Cell Adherens Junction Using Glass Micropipettes.- Using Micropatterned Supported Lipid Bilayers to Probe the Mechanosensitivity of Signaling Receptors.- Monitoring Mechano-Regulation of Gene Expression by RNA Sequencing.- Testing the Role of Focal Adhesion Kinase (FAK) in Topography-Mediated Stem Cell Differentiation by Inhibiting FAK Phosphorylation.- Long-Term Fluorescence Recovery After Photobleaching (FRAP).- Simulating 3D Cell Shape with the Cellular Potts Model.