Mechanistic understanding of how living systems sense, transduce, and respond to mechanical cues has important implications in development, physiology, and therapy. Here, the authors use an integrated atomic force microscope (AFM) and brightfield/epifluorescent microscope platform to precisely simulate living single cells or groups of cells under physiological conditions, in real time, concomitantly measuring the single-cell autophagic response and its transmission to neighboring cells. Dual-color fluorescence monitoring of the cellular autophagic response reveals the dynamics of autophagosome formation, degradation, and induction in neighboring contacting and noncontacting cells. Autophagosome formation is dependent on both the applied force and contact area of the AFM tip. More importantly, the enhancement of the autophagic responses in neighboring cells via a gap junction-dependent mechanism is observed. This AFM-based nanoacupuncture platform can serve as a tool for elucidating the primary mechanism underlying mechanical stimulation of living systems and other biomechanical therapeutics.

中文翻译：

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用原子力显微镜纳米机械诱导单细胞自噬相关荧光

对生命系统如何感知、转换和响应机械信号的机械理解对于发育、生理学和治疗具有重要意义。在这里，作者使用集成原子力显微镜（AFM）和明场/落射荧光显微镜平台实时精确模拟生理条件下的活单细胞或细胞群，同时测量单细胞自噬反应及其向邻近细胞的传输细胞。细胞自噬反应的双色荧光监测揭示了邻近接触和非接触细胞中自噬体形成、降解和诱导的动态。自噬体的形成取决于 AFM 尖端所施加的力和接触面积。更重要的是，观察到通过间隙连接依赖性机制增强邻近细胞的自噬反应。这种基于 AFM 的纳米针灸平台可以作为阐明生命系统机械刺激和其他生物力学治疗的主要机制的工具。