Simultaneous atomic force microscope (AFM) and sample scanning confocal fluorescence microscope measurements are widely used to obtain mechanistic and structural insights into protein dynamics in live cells. However, the absence of a robust technique to synchronously scan both AFM and confocal microscope piezo stages makes it difficult to visualize force-induced changes in fluorescent protein distribution in cells. To address this challenge, we have built an integrated AFM-confocal fluorescence microscope platform that implements a synchronous scanning method which eliminates image artifacts from piezo motion ramping, produces accurate pixel binning and enables the collection of a scanned image of a sample while applying force to a single point on the sample. As proof of principle, we use this instrument to monitor the redistribution of fluorescent E-cadherin, an essential transmembrane protein, in live cells, upon application of mechanical force.

中文翻译：

---

稳健的扫描同步力-荧光成像

同时原子力显微镜 (AFM) 和样品扫描共聚焦荧光显微镜测量被广泛用于获得对活细胞中蛋白质动力学的机械和结构洞察。然而，由于缺乏一种强大的技术来同步扫描 AFM 和共聚焦显微镜压电级，因此难以可视化细胞中荧光蛋白分布的力诱导变化。为了应对这一挑战，我们构建了一个集成的 AFM 共聚焦荧光显微镜平台，该平台实现了一种同步扫描方法，该方法消除了压电运动斜坡产生的图像伪影，产生准确的像素合并，并能够在施加力的同时收集样本的扫描图像样本上的一个点。作为原理证明，