Astigmatic traction force microscopy (aTFM)
Di Li, Huw Colin‐York, Liliana Barbieri, Yousef Javanmardi, Yuting Guo, Kseniya Korobchevskaya, Emad Moeendarbary, Dong Li, Marco Fritzsche
Abstract
Quantifying small, rapidly progressing three-dimensional forces generated by cells remains a major challenge towards a more complete understanding of mechanobiology. Traction force microscopy is one of the most broadly applied force probing technologies but ascertaining three-dimensional information typically necessitates slow, multi-frame z-stack acquisition with limited sensitivity. Here, by performing traction force microscopy using fast single-frame astigmatic imaging coupled with total internal reflection fluorescence microscopy we improve the temporal resolution of three-dimensional mechanical force quantification up to 10-fold compared to its related super-resolution modalities. 2.5D astigmatic traction force microscopy (aTFM) thus enables live-cell force measurements approaching physiological sensitivity.