Department of Chemistry, University of California , Berkeley, California 94720, United States.
Center for Nanomedicine and Department of Anesthesiology, Brigham and Women's Hospital, Harvard Medical School , Boston, Massachusetts 02115, United States.
Chem Rev. 2017 Jun 14;117(11):7428-7456. doi: 10.1021/acs.chemrev.6b00604. Epub 2017 Jan 3.
Correlative microscopy, the integration of two or more microscopy techniques performed on the same sample, produces results that emphasize the strengths of each technique while offsetting their individual weaknesses. Light microscopy has historically been a central method in correlative microscopy due to its widespread availability, compatibility with hydrated and live biological samples, and excellent molecular specificity through fluorescence labeling. However, conventional light microscopy can only achieve a resolution of ∼300 nm, undercutting its advantages in correlations with higher-resolution methods. The rise of super-resolution microscopy (SRM) over the past decade has drastically improved the resolution of light microscopy to ∼10 nm, thus creating exciting new opportunities and challenges for correlative microscopy. Here we review how these challenges are addressed to effectively correlate SRM with other microscopy techniques, including light microscopy, electron microscopy, cryomicroscopy, atomic force microscopy, and various forms of spectroscopy. Though we emphasize biological studies, we also discuss the application of correlative SRM to materials characterization and single-molecule reactions. Finally, we point out current limitations and discuss possible future improvements and advances. We thus demonstrate how a correlative approach adds new dimensions of information and provides new opportunities in the fast-growing field of SRM.
相关显微镜技术是指在同一样品上同时应用两种或更多种显微镜技术，其结果强调了每种技术的优势，同时弥补了各自的弱点。由于广泛的可用性、与水合和活生物样本的兼容性以及通过荧光标记实现的优异分子特异性，传统的光学显微镜在相关显微镜技术中一直是一种核心方法。然而，常规的光学显微镜的分辨率只能达到约 300nm，这削弱了其与更高分辨率方法相关联的优势。过去十年中，超分辨率显微镜（SRM）的兴起极大地提高了光学显微镜的分辨率至约 10nm，从而为相关显微镜技术带来了令人兴奋的新机遇和挑战。在这里，我们综述了如何应对这些挑战，以有效地将 SRM 与其他显微镜技术（包括光显微镜、电子显微镜、冷冻显微镜、原子力显微镜和各种形式的光谱学）相关联。虽然我们强调生物学研究，但也讨论了相关 SRM 在材料表征和单分子反应中的应用。最后，我们指出了当前的局限性，并讨论了可能的未来改进和进展。因此，我们展示了相关方法如何在快速发展的 SRM 领域中增加新的信息维度并提供新的机会。
