Zero-Mode Waveguide Nanophotonic Structures for Single Molecule Characterization.
Zero-Mode Waveguide Nanophotonic Structures for Single Molecule Characterization.
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DOI：
10.1088/1361-6463/aab8be
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发表时间：
2018-05-16
期刊：
影响因子：
--
通讯作者：
Bohn PW
中科院分区：
文献类型：
--
作者：
Crouch GM;Han D;Bohn PW
单分子表征已成为化学和生命科学中的重要研究工具，但局限性，如有限的浓度范围，无法控制分子在空间中的分布，以及内在现象，如光漂白，提出了重大挑战。非经典光学和纳米光子学的最新发展提供了有希望的途径来减轻这些限制，使得甚至可以研究低亲和力（KD ~ mM）的生物分子相互作用。在这里，我们介绍和审查特定的纳米光子设备用于支持单分子研究。光学纳米结构，如零模波导（ZMW），通常是在薄的金或铝膜制造，并用于限制光学显微光谱的观察体积为阿升至泽托升体积。这些简单的纳米结构允许分离单个分子用于光学和电化学分析，即使当感兴趣的分子以高浓度（μ M-mM）存在于本体溶液中时。ZMW阵列可以与光学探针如单分子荧光、单分子荧光共振能量转移（smFRET）和荧光相关光谱（FCS）组合，用于并行地分布式分析大量单分子反应或结合事件。此外，ZMW可以用作多功能装置，例如通过在单个离散架构中组合光学和电化学功能以实现电化学ZMW（E-ZMW）。在这篇综述中，我们将描述ZMWs的光学特性，制造和应用的单分子研究，以及ZMWs整合到系统的化学和生化分析。
Single-molecule characterization has become a crucial research tool in the chemical and life sciences, but limitations, such as limited concentration range, inability to control molecular distributions in space, and intrinsic phenomena, such as photobleaching, present significant challenges. Recent developments in non-classical optics and nanophotonics offer promising routes to mitigating these restrictions, such that even low affinity (KD ~ mM) biomolecular interactions can be studied. Here we introduce and review specific nanophotonic devices used to support single molecule studies. Optical nanostructures, such as zero-mode waveguides (ZMWs), are usually fabricated in thin gold or aluminum films and serve to confine the observation volume of optical microspectroscopy to attoliter to zeptoliter volumes. These simple nanostructures allow individual molecules to be isolated for optical and electrochemical analysis, even when the molecules of interest are present at high concentration (μM - mM) in bulk solution. Arrays of ZMWs may be combined with optical probes such as single molecule fluorescence, single molecule fluorescence resonance energy transfer (smFRET), and fluorescence correlation spectroscopy (FCS) for distributed analysis of large numbers of single-molecule reactions or binding events in parallel. Furthermore, ZMWs may be used as multifunctional devices, for example by combining optical and electrochemical functions in a single discrete architecture to achieve electrochemical ZMWs (E-ZMW). In this review, we will describe the optical properties, fabrication, and applications of ZMWs for single-molecule studies, as well as the integration of ZMWs into systems for chemical and biochemical analysis.
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