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Protocol for measuring weak cellular traction forces using well-controlled ultra-soft polyacrylamide gels
Farah Mustapha1,2,3,4,5, Kheya Sengupta4,5, Pierre-Henri Puech1,2,3,4
1Laboratoire Adhésion et Inflammation (LAI), Aix Marseille University, LAI UM 61, Marseille 13288, France.
STAR Protocols
|February 7, 2022
Summary
This study presents an easy in vitro traction force microscopy (TFM) protocol using ultrasoft gels for detecting weak cellular stresses. The method simplifies quantifying traction forces, demonstrated with peripheral blood mononuclear cells.
Area of Science:
- Biophysics
- Cell Biology
- Biomaterials
Background:
- Traction Force Microscopy (TFM) is crucial for analyzing cellular forces.
- Fabricating ultrasoft substrates for detecting low cellular stresses (<100 Pa) is challenging.
Purpose of the Study:
- To develop a simple in vitro TFM protocol for reproducible ultrasoft substrate fabrication.
- To enable reliable quantification of weak cellular stresses.
Main Methods:
- Utilized protein-coated polyacrylamide gels as ultrasoft substrates.
- Employed wide-field fluorescence microscopy for imaging.
- Developed open-source and in-house scripts for data analysis.
Main Results:
- Successfully fabricated reproducible ultrasoft substrates for TFM.
- Quantified traction stresses below 100 Pa.
- Demonstrated the protocol using peripheral blood mononuclear cells.
Conclusions:
- The developed TFM protocol offers a simplified approach for studying cellular mechanics.
- Enables accurate measurement of weak cellular stresses using accessible methods.
