STORM和STED显微成像技术特点的比较

Comparison of the characteristics of STORM and STED micro-imaging techniques

随机光学重建显微镜(stochastic optical reconstruction microscopy,STORM)技术和受激发射损耗(stimulated emission depletion,STED)显微镜技术是近年来发展迅速的两种超分辨率荧光显微镜技术。这两种技术均提供超越传统荧光显微镜分辨率成像的功能,具有多色显像,三维成像以及活细胞内成像的潜力。在这篇综述中,我们关注两种技术荧光控制、激光强度等技术参数设定,同时结合样品制备、图像采集与处理等流程优化对比两者在分辨率、图像采集时间及具体应用中的优劣。STORM可获得更高的三维分辨率,但可能需要更长的图像采集时间。STED需要较高损耗光强度,却能在图像采集后立即生成超分辨率图像,不需要额外图像数据处理。最终,选择STORM和STED不仅取决于技术的具体应用,还取决于操作者优化各环节技术参数的能力,从而决定图像质量。

Stochastic optical reconstruction microscopy (STORM) and stimulated emission depletion (STED) microscopy are two super-resolution fluorescence microscopic technologies developed in recent years. Both technologies can transcend traditional fluorescence microscopy image resolution, with the potential for multiple colors, three-dimensional imaging, and live-cell intracellular imaging. In this review, we focus on fluorescence control, laser intensity and other technical parameters, as well as sample preparation, image acquisition and processing optimization of the two processes, and compare their strengths and weaknesses in resolution, image acquisition time, and specific applications. Overall, STORM provides higher 3D-resolution, but may require longer image acquisition times. STED requires higher laser intensity, but can generate images immediately after image acquisition, without additional image data processing. Ultimately, choosing STORM or STED depends not only on the specific application of the technology, but also on the ability of the operator to optimize the technical parameters at each step.