Bates Mark, Jones Sara A, Zhuang Xiaowei
Cold Spring Harb Protoc. 2013 Jun 1;2013(6):540-1. doi: 10.1101/pdb.prot075168.
Stochastic optical reconstruction microscopy (STORM) is a method for superresolution fluorescence imaging based on the high accuracy localization of individual fluorophores. It uses optically switchable fluorophores: molecules that can be switched between a nonfluorescent and a fluorescent state by exposure to light. Many synthetic fluorescent dye molecules show photoswitchable fluorescence emission. In particular, photoswitchable cyanine fluorophores such as Cy5, Alexa 647, and Cy7, may be paired with a second fluorophore, which serves as an activator, determining the wavelength of light that re-activates the fluorescence of the photoswitchable molecule. This protocol describes the preparation of antibodies labeled with one such pairing scheme of synthetic fluorophores, Alexa 405 and Alexa 647. It may easily be adapted for labeling with other fluorophores or for labeling other substrate molecules.
随机光学重建显微镜（STORM）是一种基于单个荧光团高精度定位的超分辨率荧光成像方法。它使用可光开关荧光团：即通过光照可在非荧光状态和荧光状态之间切换的分子。许多合成荧光染料分子表现出光开关荧光发射。特别是，诸如Cy5、Alexa 647和Cy7等可光开关花菁荧光团可与第二种荧光团配对，该荧光团用作激活剂，决定重新激活可光开关分子荧光的光的波长。本方案描述了用一种这样的合成荧光团（Alexa 405和Alexa 647）配对方案标记抗体的制备方法。它可以很容易地适用于用其他荧光团标记或标记其他底物分子。
