Quantifying cell generated mechanical forces is key to furthering our understanding of mechanobiology. Traction force microscopy (TFM) is one of the most broadly applied force probing technologies, but its sensitivity is strictly dependent on the spatio-temporal resolution of the underlying imaging system. In previous works, it was demonstrated that increased sampling densities of cell derived forces permitted by super-resolution fluorescence imaging enhanced the sensitivity of the TFM method. However, these recent advances to TFM based on super-resolution techniques were limited to slow acquisition speeds and high illumination powers. Here, we present three novel TFM approaches that, in combination with total internal reflection, structured illumination microscopy and astigmatism, improve the spatial and temporal performance in either two-dimensional or three-dimensional mechanical force quantification, while maintaining low illumination powers. These three techniques can be straightforwardly implemented on a single optical set-up offering a powerful platform to provide new insights into the physiological force generation in a wide range of biological studies.

This article is part of the Theo Murphy meeting issue ‘Super-resolution structured illumination microscopy (part 1)'.

量化细胞产生的机械力是进一步了解机械生物学的关键。牵引力显微镜 (TFM) 是应用最广泛的力探测技术之一，但其灵敏度严格取决于基础成像系统的时空分辨率。在以前的工作中，已经证明超分辨率荧光成像允许增加的细胞衍生力的采样密度提高了 TFM 方法的灵敏度。然而，这些基于超分辨率技术的 TFM 的最新进展仅限于低采集速度和高照明功率。在这里，我们提出了三种新颖的 TFM 方法，结合全内反射、结构化照明显微镜和散光，改善二维或三维机械力量化的空间和时间性能，同时保持低照明功率。这三种技术可以直接在单个光学装置上实施，提供了一个强大的平台，为广泛的生物学研究中的生理力产生提供了新的见解。

本文是 Theo Murphy 会议问题“超分辨率结构照明显微镜（第 1 部分）”的一部分。