Field Guide to Traction Force Microscopy

Area of Science:

• Biophysics
• Cell Biology
• Biomaterials Science

Background:

• Traction Force Microscopy (TFM) is crucial for measuring cell contractility on biomimetic substrates.
• Many TFM workflows involve user choices with unclear impacts on quantitative results.
• Limited publications detail TFM's complete experimental and mathematical steps, hindering understanding of decision consequences.

Purpose of the Study:

• To provide a "Field Guide" explaining the mathematical basis of common TFM methods.
• To focus on error propagation in TFM workflows based on experimental and analytical choices.
• To empower experimentalists to quantify cell contractility with confidence.

Main Methods:

• Conceptual review and analysis of TFM research over the last two decades.
• Explanation of mathematical underpinnings for common TFM techniques.
• Focus on error propagation linked to specific experimental design and analytical decisions.

Main Results:

• Covers substrate manufacturing, mechanical properties, imaging, and image processing.
• Details methods for calculating traction stress and data reporting strategies.
• Highlights assumptions and considerations critical for TFM workflows.

Conclusions:

• Offers researchers a better understanding of TFM options for diverse cell types.
• Facilitates informed decision-making in TFM workflow design.
• Aims to enhance confidence in quantitative cell contractility measurements.