Extended super-resolution three-dimensional mechanical probing in living cells.

机械生物学的新视角目前正在物理和生物医学研究领域的多个学科中出现。与传统观点相反，最近的证据表明，细胞不仅在外部刺激引发的信号事件的下游调节其细胞力学，而且细胞采用其细胞骨架的多种反馈机制，使其能够动态地调节细胞力学以满足生理需要。因此，这提供了一个以前无法预见的画面，其中细胞主动施加和抵抗生物力学力，以调整其机械生物学，从而促进其功能。因此，在生物工程和生物医学科学的界面上，量化细胞力已经成为跨多个学科的重要使命。该项目的总体目标是开发新一代力探测方法，以前所未有的精度和分辨率在活细胞中进行生理机械探测。为了设计这项技术，我们将联合收割机结合最先进的3D高速全内反射（TIRF）和尖端的超分辨率结构照明（SIM）显微镜的新技术，再加上扩展的机械力TFM探测，以创造一种新的突破性技术，用于活细胞中的机械探测。我们将通过量化各种粘附细胞中的机械力产生和激活免疫T细胞来展示新方法的力量。我们预计，我们的研究可能会导致取代传统的TFM测量与细胞机械生物学的理解的重大影响。最终，我们的目标是将3D eTIRF-SIM-TFM方法商业化。

New perspective of mechanobiology is currently emerging across multiple disciplines in the physical and biomedical research fields. In contrast to conventional beliefs, recent evidence indicates that cells regulate their cell mechanics not only downstream of signalling events triggered by external stimuli, but that cells employ a diversity of feedback mechanisms of their cytoskeleton enabling them to dynamically adjust cell mechanics to meet physiological needs. Consequently, this provides a previously unforeseen picture wherein cells actively exert and resist biomechanical force to tune their mechanobiology, and thus facilitate their function. Quantifying cellular forces has therefore become an important mission across multiple disciplines at the interface of bioengineering and biomedical sciences. The overall goal of this project is the development of a new-generation force probing methodology to enable physiological mechanical probing in living cells at unprecedented accuracy and resolution. To engineer this technology, we will combine a new technique of state-of-the-art 3D high-speed total-internal-reflection (TIRF) and cutting-edge super-resolution structural-illumination (SIM) microscopy coupled with extended mechanical force TFM probing to create a novel breakthrough technology for mechanical probing in living cells. We will demonstrate the power of the new methodology by quantifying mechanical force production in a variety of adherent cells and activating immune T cells. We anticipate that our research might lead to the replacement of conventional TFM measurements with major implications for the understanding of the cellular mechanobiology. Ultimately, we therefore aim to commercialise the 3D eTIRF-SIM-TFM method.