Mechanobiology: Methods and Protocols

ISBN-13: 9781071628508 / Angielski / Twarda / 2023 / 343 str.

Ronen Zaidel-Bar

Opis

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. 

Authoritative and practical, Mechanobiology: Methods and Protocols aims to provide meaningful tools for cell and developmental biologists approaching the study of cell and tissue dynamics from a mechanobiological perspective, molecular biologists interested in the effects of force on proteins, as well as for cancer biologists and biophysicists.

Spis treści

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-Muñoz

 

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 Püllen, 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 Chardès, Raphaël Clément and Pierre-François 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 Chabouté, 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 Windgasse and Carsten Grashoff

 

16. Visualizing Neurons under Tension In Vivo with Optogenetic Molecular Force Sensors

            Neus Sanfeliu-Cerdán, Li-Chun Lin, Alexander R. Dunn, Miriam B. Goodman, and Michael Krieg

 

17. Single-Cell Quantification of the Mechanical Stability of Cell-Cell Adherens Junction Using Glass Micropipettes

            Wenmao Huang, Chaoyu Fu, and Jie Yan

 

Part VI: Probing Cell Signaling and Gene Expression in Response to Force

 

18. Using Micropatterned Supported Lipid Bilayers to Probe the Mechanosensitivity of Signaling Receptors

            Zhongwen Chen

 

19. Monitoring Mechano-Regulation of Gene Expression by RNA Sequencing

            Yekaterina A. Miroshnikova

 

20. Testing the Role of Focal Adhesion Kinase (FAK) in Topography-Mediated Stem Cell Differentiation by Inhibiting FAK Phosphorylation

            Yuan Yao and Evelyn K.F. Yim

 

Part VII: Biophysical Modeling of Cell Shape and Protein Dynamics

 

21. Long-Term Fluorescence Recovery After Photobleaching (FRAP)

            Takumi Saito, Daiki Matsunaga, and Shinji Deguchi

 

22. Simulating 3D Cell Shape with the Cellular Potts Model

            Rabea Link and Ulrich S. Schwarz