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A Biologist's Guide to Traction Force Microscopy Using Polydimethylsiloxane Substrate for Two-Dimensional Cell
Jessica L Teo1, Chwee Teck Lim2,3,4, Alpha S Yap1
1Division of Cell and Developmental Biology, Institute for Molecular Bioscience, The University of Queensland, St. Lucia, Brisbane, Queensland 4072, Australia.
STAR Protocols
|October 28, 2020
Summary
This study presents a simple in vitro traction force microscopy (TFM) protocol for measuring cellular forces in epithelial cells. The method uses readily available materials and detailed analysis, aiding mechanobiology research.
Area of Science:
- Biophysics
- Cell Biology
- Biomaterials Science
Background:
- Cellular traction forces are critical for epithelial tissue dynamics, influencing processes like wound healing and cell extrusion.
- Understanding these forces requires robust experimental methodologies.
Purpose of the Study:
- To describe a straightforward in vitro protocol for Traction Force Microscopy (TFM).
- To enable researchers to quantify cellular traction forces in epithelial cells for mechanobiology studies.
- To provide guidance on selecting appropriate TFM protocols based on experimental needs.
Main Methods:
- Utilized polydimethylsiloxane (PDMS) substrates coated with extracellular matrix (ECM) proteins.
- Employed widefield fluorescence microscopy for image acquisition.
- Included detailed analytical steps for calculating traction forces.
Main Results:
- Successfully established a simple and accessible TFM protocol.
- Demonstrated the application of the protocol for studying epithelial cell mechanobiology.
- Provided comparative insights into TFM protocols using different substrate types.
Conclusions:
- The described TFM protocol offers a practical approach for investigating cellular forces in epithelial systems.
- This method facilitates the study of mechanobiology, contributing to a deeper understanding of epithelial cell functions.
- Guidelines are provided for choosing between PDMS-based and polyacrylamide hydrogel-based TFM methods.
