INM-Leibniz-Institute for New Materials, Nano Cell Interactions Group, Saarbrücken, Germany.
Chemphyschem. 2012 Jun 4;13(8):1986-2000. doi: 10.1002/cphc.201100986. Epub 2012 Feb 28.
For about a decade, superresolution fluorescence microscopy has been advancing steadily, maturing from the proof-of-principle stage to routine application. Of the various techniques, STED (stimulated emission depletion) microscopy was the first to break the diffraction barrier. Today, it is a prominent and versatile form of superresolution light microscopy. STED microscopy has shed a sharper light on numerous topics in cell biology, but also in material sciences. Both disciplines extend into the nanometer range, making detailed studies of structural and functional relationships difficult or even impossible to achieve using diffraction-limited microscopy. With recent advancements like spectral multiplexing or live-cell imaging, STED microscopy makes nanoscale materials and components of the cell accessible for fluorescence-based investigations. With multicolor superresolution imaging, even the interactions between biological and engineered nanostructures can be studied in detail. This review gives an introduction into the working principle of STED microscopy, provides a detailed overview of recent advancements and new techniques implemented for use with STED microscopy and shows how these have been applied in the life sciences and nanotechnologies.
在过去的十年中，超分辨率荧光显微镜技术稳步发展，从原理验证阶段成熟到常规应用。在各种技术中，受激发射损耗（STED）显微镜是第一个突破衍射障碍的技术。如今，它是一种出色且多功能的超分辨率光学显微镜形式。STED 显微镜不仅在细胞生物学领域，而且在材料科学领域，都为众多主题提供了更清晰的认识。这两个学科都延伸到纳米范围，使得使用衍射受限显微镜难以甚至不可能对结构和功能关系进行详细研究。随着最近的进展，如光谱复用或活细胞成像，STED 显微镜使得荧光基研究能够接触到纳米级的材料和细胞成分。通过多色超分辨率成像，甚至可以详细研究生物和工程纳米结构之间的相互作用。本文综述了 STED 显微镜的工作原理，详细介绍了最近的进展和为 STED 显微镜实施的新技术，并展示了它们在生命科学和纳米技术中的应用。
