Scanning Resonator Microscopy

总结 光激活定位显微镜（PALM）和随机光学重建显微镜 （风暴）通过引入合理的方法，彻底改变了超分辨率领域 在最终用户实验室中掌握和实施。它们的影响现在已经渗透到各个学科， 它们的独特能力正被用于解决日益广泛的重要问题。 近场扫描光学显微镜（NSOM）是另一种超分辨率技术， 与PALM和STORM的属性互补。NSOM是一种扫描探针技术， 专门制造的光纤探针，用于测量超分辨率荧光和样品形貌， 同步这在生物科学中特别有用，其中细胞形状和形态 特征可以直接与荧光图像中特别标记的物质进行比较。到目前为止， 然而，NSOM在生物科学中的影响是有限的。这主要是由于 繁琐的实施要求和光纤探头的不良性能。克服 这些挑战，我们提出了一种全新的方法，将光学对比度机制与 原子力显微镜（AFM） 扫描共振显微镜（SRM）使用一个小的电介质微球连接在末端， 用于超分辨率成像的常规AFM探针。该方法利用了回音壁模式 (WGM)在所附接的谐振器中激发谐振以感测或激发样品特性。不像 传统的NSOM，探针很容易在解剖显微镜下组装，SRM需要 只有最小的修改商业AFM平台。这应该能够在世界范围内广泛采用。 终端用户实验室我们已经开发了一个原型SRM，并证明了这种方法的可行性， 同时定量样品的折射率和超分辨率薄膜的形貌。的 这里的总体目标是开发下一代SRM，能够同时荧光，折射 复杂生物样品的指数和地形测量。 本文提出的研究计划包括：（目标1）研制一种用于固体火箭发动机尖端的“沿沿着”式光纤耦合器 激发，使超分辨率荧光成像厚生物样品和（目标2）测试， 通过研究膜联蛋白VI的定位，在真实的生物系统上验证和基准SRM性能 在钙刺激后固定的人动脉平滑肌细胞中。成功完成本 工作将介绍一种新的超分辨率工具，可以很容易地在最终用户实验室采用，具有独特的 与现有技术互补。对于未来，很容易设想额外的传感器 通过特定的涂层或光学谐振腔的修改， 尖端。

Summary Photoactivated localization microscopy (PALM) and stochastic optical reconstruction microscopy (STORM) have revolutionized the super-resolution field by introducing approaches that are reasonably mastered and implemented in the end-user laboratory. Their impact is now felt across the disciplines where their unique capabilities are being applied to an ever-expanding spectrum of important problems. Near-field scanning optical microscopy (NSOM) is another super-resolution technique that has attributes complementary to those of PALM and STORM. NSOM is a scanning probe technique that uses specially fabricated fiber optic probes to measure super-resolution fluorescence and sample topography, simultaneously. This is particularly useful in the biological sciences, where cell shape and morphological features can be compared directly with species specifically labeled in the fluorescence image. To date, however, the impact of NSOM in the biological sciences has been modest. This is mainly due to burdensome implementation requirements and poor performance of the fiber optic probes. To overcome these challenges, we propose a completely new approach for integrating optical contrast mechanisms with atomic force microscopy (AFM). Scanning resonator microscopy (SRM) uses a small dielectric microsphere attached at the end of conventional AFM probe for super-resolution imaging. The approach exploits whispering gallery mode (WGM) resonances excited in the attached resonator to sense or excite sample properties. Unlike conventional NSOM, the probes are easily assembled under a dissecting microscope and SRM requires only minimal modifications to commercial AFM platforms. This should enable widespread adoption in the end user lab. We have developed a prototype SRM and demonstrated the feasibility of this approach by simultaneously quantifying sample refractive index and topography of thin films with super-resolution. The overall goal here is to develop the next generation SRM capable of simultaneous fluorescence, refractive index, and topography measurements on complex biological samples. The research plan proposed here will: (Aim 1) develop a “ride along” fiber optic coupler for SRM tip excitation that enables super-resolution fluorescence imaging on thick biological samples and (Aim 2) test, validate, and benchmark SRM performance on a real biological system by studying annexin VI localization in fixed human arterial smooth muscle cells following calcium stimulation. The successful completion of this work will introduce a new super-resolution tool that can easily be adopted in the end user lab, with unique capabilities complementary to existing technologies. For the future, it is easy to envision additional sensing capabilities being integrated with SRM through specific coatings or modifications of the optical resonator at the tip end.