利用对比增强显微镜对非常小的单个纳米颗粒进行高速成像和跟踪。

High-speed imaging and tracking of very small single nanoparticles by contrast enhanced microscopy.

机构信息

Institute of Atomic and Molecular Sciences (IAMS), Academia Sinica, 10617 Taipei, Taiwan.

出版信息

Nanoscale. 2019 Jan 3;11(2):568-577. doi: 10.1039/c8nr06789a.

Abstract

Nanoparticles have been used extensively in biology-related research and many applications require direct visualization of individual nanoparticles by optical microscopy. For long-term and high-speed measurements, scattering-based microscopy is a unique technique because of the stable and indefinite scattering signals. In scattering-based single-particle measurements, large nanoparticles are usually needed in order to generate sufficient signals for detection. However, larger nanoparticles introduce greater mass loading, experience stronger steric hindrance, and are more prone to crosslinking. In this work, we demonstrate coherent brightfield (COBRI) microscopy with enhanced contrast and show its capability of direct visualization of very small nanoparticles in scattering at a high speed. COBRI microscopy allows us to visualize and track single metallic and dielectric nanoparticles, as small as 10 nm, at 1000 frames per second. A quantitative relationship between the linear scattering cross section of the nanoparticle and its COBRI contrast is reported. Using COBRI microscopy, we further demonstrate the tracking of 10 nm gold nanoparticles labeled to lipid molecules in supported bilayer membranes, showing that the small nanoparticles may facilitate single-molecule measurements with reduced perturbation. Furthermore, the identical imaging sensitivities of COBRI and interferometric scattering (iSCAT) microscopy, the reflection counterpart of COBRI, is demonstrated at an equal illumination intensity. Finally, future improvements in the speed and sensitivity of scattering-based interference microscopy are discussed.

摘要

纳米粒子在生物学相关研究中得到了广泛应用,许多应用需要通过光学显微镜直接观察单个纳米粒子。对于长期和高速测量,基于散射的显微镜是一种独特的技术,因为其散射信号稳定且无期限。在基于散射的单粒子测量中,通常需要较大的纳米粒子才能产生足够的检测信号。然而,较大的纳米粒子会引入更大的质量负载,经历更强的空间位阻,并且更容易交联。在这项工作中,我们展示了具有增强对比度的相干明场(COBRI)显微镜,并展示了其在高速散射中直接可视化非常小的纳米粒子的能力。COBRI 显微镜使我们能够以每秒 1000 帧的速度可视化和跟踪单个金属和介电纳米粒子,小至 10nm。报告了纳米粒子的线性散射截面与其 COBRI 对比度之间的定量关系。使用 COBRI 显微镜,我们进一步展示了在支撑双层膜中标记脂质分子的 10nm 金纳米粒子的跟踪,表明小纳米粒子可能有助于减少干扰的单分子测量。此外,还证明了 COBRI 的反射对应物干涉散射(iSCAT)显微镜在相同的照明强度下具有相同的成像灵敏度。最后,讨论了散射干涉显微镜在速度和灵敏度方面的未来改进。

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