Two-color far-field fluorescence nanoscopy based on photoswitchable emitters
Two-color far-field fluorescence nanoscopy based on photoswitchable emitters
复制标题
DOI：
10.1007/s00340-007-2729-0
复制
发表时间：
2007-07-01
影响因子：
2.1
通讯作者：
Eggeling, C.
中科院分区：
文献类型：
--
作者：
Bock, H.;Geisler, C.;Eggeling, C.
我们展示了基于单个荧光标记的光开关的具有纳米级空间分辨率的双色远场荧光显微镜。通过启用、记录和禁用可逆可切换荧光蛋白rsFastLime和有机荧光团花青5的发射，我们记录了整个细胞内的双色纳米级图像。单个发射器的位置以20 nm的典型精度确定，这在很大程度上构成了系统的横向分辨率。双色共定位实验中的光开关是远场荧光纳米显微镜在大分子水平上研究（生物）样品的重要一步。
We demonstrate two-color far-field fluorescence microscopy with nanoscale spatial resolution based on the photoswitching of individual fluorescent markers. By enabling, recording, and disabling the emission of the reversibly switchable fluorescent protein rsFastLime and of the organic fluorophore cyanine5, we recorded two-color nanoscale images inside whole cells. The position of individual emitters was determined with a typical accuracy of 20 nm, which largely constitutes the lateral resolution of the system. Photoswitching in two-color colocalization experiments represents a major step towards the application of far-field fluorescence nanoscopy to the study of (biological) samples on the macromolecular level.