Guidance for 3D traction force microscopy today and in the next decade

Area of Science:

• Mechanobiology
• Cellular mechanics
• Biophysics

Background:

• Mechanobiology investigates how mechanical forces impact cellular functions.
• Traction Force Microscopy (TFM) is a key technique for measuring cell-exerted forces on the extracellular matrix.
• Three-dimensional TFM (3DTFM) has advanced but remains underutilized compared to 2D TFM.

Purpose of the Study:

• To address common skepticism surrounding the utility of 3DTFM.
• To detail experimental and computational strategies for overcoming 3DTFM limitations.
• To guide the broader adoption and future development of 3DTFM in mechanobiology.

Main Methods:

• Review of current experimental and computational approaches for 3DTFM.
• Integration strategies of 3DTFM with biological readouts for long-term studies.
• Discussion of data interpretation metrics and traction recovery methods.

Main Results:

• Identified and addressed key limitations and skepticism associated with 3DTFM.
• Provided methods for integrating 3DTFM with biological readouts and long-term experiments.
• Outlined strategies for data interpretation and application to specific biological questions.

Conclusions:

• 3DTFM has significant potential for advancing mechanobiology research.
• Addressing current limitations and promoting standardized methods will increase 3DTFM adoption.
• Future directions include combining 3DTFM with emerging technologies for complex 3D cellular analyses.