同步、无串扰相关AFM和共聚焦显微镜/光谱
Synchronous, Crosstalk-free Correlative AFM and Confocal Microscopies/Spectroscopies
摘要：
显微镜已成为我们观察生物系统和组件的表征工具的支柱。不同显微镜的相关和同步使用依赖于图像采集过程中无干扰的基本假设。在这项工作中，通过探索原子力显微镜和共焦荧光寿命成像显微镜（AFM-FLIM）的相关使用，我们量化同步采集过程中发生的串扰效应。我们的特点和最小化的光机械力在不同的原子力显微镜的杠杆干扰正常的原子力显微镜操作，以及假发光的尖端和悬臂影响时间分辨荧光检测。通过定义非干扰实验成像参数，我们显示了准确的实时采集和相互作用力，荧光寿命和强度表征形态（AFM）和局部粘度（FLIM）的凝胶和流体相分离的支持脂质模型膜的二维映射。最后，作为通过同步力和荧光光谱的原理证明，我们通过控制其与金属表面的距离来精确地调整位于AFM针尖上的荧光纳米金刚石的寿命。这开辟了一个新的途径淬火传感图像软生物样品，如膜，因为它不需要与传统的AFM在液体中的尖端样品的机械接触。
英文摘要：
Microscopies have become pillars of our characterization tools to observe biological systems and assemblies. Correlative and synchronous use of different microscopies relies on the fundamental assumption of non-interference during images acquisitions. In this work, by exploring the correlative use of Atomic Force Microscopy and confocal-Fluorescence-Lifetime Imaging Microscopy (AFM-FLIM), we quantify cross-talk effects occurring during synchronous acquisition. We characterize and minimize optomechanical forces on different AFM cantilevers interfering with normal AFM operation as well as spurious luminescence from the tip and cantilever affecting time-resolved fluorescence detection. By defining non-interfering experimental imaging parameters, we show accurate real-time acquisition and two-dimensional mapping of interaction force, fluorescence lifetime and intensity characterizing morphology (AFM) and local viscosity (FLIM) of gel and fluid phases separation of supported lipid model membranes. Finally, as proof of principle by means of synchronous force and fluorescence spectroscopies, we precisely tune the lifetime of a fluorescent nanodiamond positioned on the AFM tip by controlling its distance from a metallic surface. This opens up a novel pathway of quench sensing to image soft biological samples such as membranes since it does not require tip-sample mechanical contact in contrast with conventional AFM in liquid.
机构、总结、中文摘要均由AI分析而来，可能存在错误，请以论文正文为准。
