Mechanical Fidelity of Fixed-Cell Traction Force Microscopy

Glutaraldehyde fixation in traction force microscopy preserves up to 99% of displacement and traction magnitudes and strain energy compared with the same live cells.

• Why it matters: It is unclear whether fixed-cell TFM reproduces the live-cell mechanical state, which limits reliable use of post-fixation fluorescence workflows.
• What they did: The same cells were measured before and after fixation using different fixation protocols and substrate stiffnesses, then displacement, traction, and strain-energy fields were compared.
• The result: Fixation largely maintained the spatial organization and direction of substrate displacements and reconstructed traction fields, while magnitudes changed by protocol; GA showed the closest live-cell agreement, enabling practical multi-well fixed-cell TFM with fluorescence imaging.

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