Kuang Cuifang, Zhao Wei, Wang Guiren
Department of Mechanical Engineering and Biomedical Engineering Program, University of South Carolina, Columbia, South Carolina 29208, USA.
Rev Sci Instrum. 2010 May;81(5):053709. doi: 10.1063/1.3432001.
Stimulated emission depletion (STED) microscopy is one of the breakthrough technologies that belong to far-field optical microscopy and can achieve nanoscale spatial resolution. We demonstrate a far-field optical nanoscopy based on continuous wave lasers with different wavelengths, i.e., violet and green lasers for excitation and STED, respectively. Fluorescent dyes Coumarin 102 and Atto 390 are used for validating the depletion efficiency. Fluorescent nanoparticles are selected for characterizing the spatial resolution of the STED system. Linear scanning of the laser beams of the STED system along one line of a microscope slide, which is coated with the nanoparticles, indicates that a spatial resolution of about 70 nm has so far been achieved. A two-dimensional image of the particle pattern of the STED system is constructed and compared with scanning confocal microscope. The present work has further extended the application of the STED microscopy into the blue regime.
受激发射损耗（STED）显微镜是远场光学显微镜中的突破性技术之一，能够实现纳米级空间分辨率。我们展示了一种基于不同波长连续波激光器的远场光学纳米显微镜，即分别使用紫光激光器和绿光激光器进行激发和受激发射损耗。荧光染料香豆素102和Atto 390用于验证损耗效率。选择荧光纳米颗粒来表征STED系统的空间分辨率。STED系统的激光束沿着涂有纳米颗粒的显微镜载玻片的一条线进行线性扫描，结果表明目前已实现约70纳米的空间分辨率。构建了STED系统颗粒图案的二维图像，并与扫描共聚焦显微镜进行了比较。本工作进一步将STED显微镜的应用扩展到了蓝光领域。
