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Cellular traction forces (CTFs) play an integral role in both physiological processes and disease, and are a topic of interest in mechanobiology. Traction force microscopy (TFM) is a family of methods used to quantify CTFs in a variety of settings. State-of-the-art 3D TFM methods typically rely on confocal fluorescence microscopy, which can impose limitations on acquisition speed, volumetric coverage, and temporal sampling or coverage. In this report, we present the first quantitative implementation of a new TFM technique: traction force optical coherence microscopy (TF-OCM). TF-OCM leverages the capabilities of optical coherence microscopy and computational adaptive optics (CAO) to enable the quantitative reconstruction of 3D CTFs in scattering media with minute-scale temporal sampling. We applied TF-OCM to quantify CTFs exerted by isolated NIH-3T3 fibroblasts embedded in Matrigel, with five-minute temporal sampling, using images spanning a 500 × 500 × 500 μm 3  field-of-view. Due to the reliance of TF-OCM on computational imaging methods, we have provided extensive discussion of the equations, assumptions, and failure modes of these methods. By providing high-throughput, label-free, volumetric imaging in scattering media, TF-OCM is well-suited to the study of 3D CTF dynamics, and may prove advantageous for the study of large cell collectives, such as the spheroid models prevalent in mechanobiology.

Quantitative reconstruction of time-varying 3D cell forces with traction force optical coherence microscopy