Single molecule localization microscopy for superresolution
Single molecule localization microscopy for superresolution
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DOI：
10.1088/2040-8978/15/9/094001
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发表时间：
2013-09-01
影响因子：
2.1
通讯作者：
Davidson, Michael W.
中科院分区：
文献类型：
--
作者：
Allen, John R.;Ross, Stephen T.;Davidson, Michael W.
近年来，光学显微镜取得了突飞猛进的发展，绕过了与样品和探测器之间发生的光的衍射有关的传统分辨率限制。这些技术统称为“超分辨率”显微镜。超分辨技术的一个主要家族，可变地被称为Palm，FPALM和STORM，使用对荧光团的激发状态的时间控制来顺序地识别时间和空间上的单个非重叠发射体。单点发射体的常规图像被拟合到次衍射限制区域，并根据在数千个单独成像帧上收集的位置数据重建合成图像。本文简要概述了超分辨率显微镜，然后详细讨论了STORM，包括样品制备的实用指南，旨在帮助非专业人员更容易地使用这项技术。
In recent years there has been a rash of developments in light microscopies circumventing traditional resolution limits associated with the diffraction of light occurring between the sample and the detector. Collectively, these techniques are referred to as 'superresolution' microscopies. One major family of superresolution techniques, variably referred to as PALM, FPALM and STORM, uses temporal control of the excited state of fluorophores to sequentially identify single non-overlapping emitters in time and space. Conventional images of single point emitters are fitted to sub-diffraction-limited areas, and a composite image is reconstructed from position data collected over many thousands of individual imaging frames. This paper provides a brief overview of superresolution microscopy, followed by a detailed discussion of STORM, including practical guidelines for sample preparation designed to help to make the technique more accessible to the non-specialist.