Mechanobiology: Methods and Protocols

Part I: Quantifying the Mechanical Properties of Cells and Adhesion Proteins

1. Measuring Cell Mechanical Properties Using Microindentation

Julien Husson

2. In Situ Measurements of Cell Mechanical Properties Using Force-Spectroscopy

Meital Reches

3. Quantification of Apparent Membrane Tension and Membrane-to-Cortex Attachment in Animal Cells Using Atomic Force Microscopy-Based Force Spectroscopy

Martin Bergert and Alba Diz-Muoz

4. Characterizing the Biophysical Properties of Adhesive Proteins in Live Cells Using Single Molecule Atomic Force Microscopy

Andrew Vae Priest and Sanjeevi Sivasankar

Part II: Exposing Cells to External Mechanical Forces

5. Application of Shear Stress to Endothelial Cells Using a Parallel Plate Flow Chamber

Jason Sedlak and Alisa Clyne

6. Cell Stretcher Assay to Analyze Mechanoresponses to Cyclic Stretching

Robin Pllen, Jens Konrad, Bernd Hoffmann, and Rudolf Merkel

7. Two-Point Optical Manipulation of Cell Junctions in the Early Epithelium of the Drosophila Embryo

Kenji Nishizawa, Claire Chards, Raphal Clment and Pierre-Franois Lenne

Part III: Quantitative Characterization of Cell Mechanosensing

8. A Microfluidic-Like System (MLS) to Grow, Image, and Quantitatively Characterize Rigidity-Sensing by Plant's Roots and Root Hair Cells

David Pereira, Thomas Alline, Gaurav Singh, Marie-Edith Chabout, and Atef Asnacios

9. Photoresponsive Hydrogels for Studying Mechanotransduction of Cells

Keun-Young Park, David J. Odde, and Mark D. Distefano

10. Analyzing Mechanically Evoked Currents at Cell-Substrate Junctions

Surabhi Shrestha, Jessica Richardson, and Kate Poole

11. Quantifying Strain Sensing Protein Recruitment During Stress Fiber Repair

Shailaja Seetharaman, Stefano Sala, Margaret L. Gardel, and Patrick W. Oakes

Part IV: Measuring Forces Applied by Cells on the Extracellular Matrix

12. Quantification of Invadopodia Formation and Matrix Degradation Activity

Jubina Balan Venghateri and Benjamin Geiger

13. Measuring Cellular Traction Forces with Micropillar Arrays

Malak Amer and Haguy Wolfenson

14. Imaging Cell Adhesive Force at the Single Molecule Level

Ying Tu and Xuefeng Wang

Part V: Force Measurement Inside Cells

15. Multiplexed Molecular Tension Sensor Measurements Using PIE-FLIM

Lukas Win