Nanometer resolution imaging and tracking of fluorescent molecules with minimal photon fluxes
Nanometer resolution imaging and tracking of fluorescent molecules with minimal photon fluxes
复制标题
DOI：
10.1126/science.aak9913
复制
发表时间：
2017-02-10
期刊：
影响因子：
56.9
通讯作者：
Hell, Stefan W.
中科院分区：
文献类型：
--
作者：
Balzarotti, Francisco;Eilers, Yvan;Hell, Stefan W.
我们介绍MINFLUX，在空间中定位光子发射器的概念。通过用激发光的局部强度最小值探测发射器，MINFLUX最大限度地减少了高定位精度所需的荧光光子。在我们的实验中，与流行的质心定位相比，需要少22倍的荧光光子。在超分辨显微镜中，MINFLUX达到了类似于1纳米的精度，分辨分子的距离只有6纳米。MINFLUX跟踪单个荧光蛋白的时间分辨率和定位的数量增加了100倍，如在活大肠杆菌中扩散30S核糖体亚基所示。由于尚未达到概念上的限制，我们希望这种定位方式为观察活细胞及其他大分子的动态，分布和结构开辟新的天地。
We introduce MINFLUX, a concept for localizing photon emitters in space. By probing the emitter with a local intensity minimum of excitation light, MINFLUX minimizes the fluorescence photons needed for high localization precision. In our experiments, 22 times fewer fluorescence photons are required as compared to popular centroid localization. In superresolutionmicroscopy, MINFLUXattained similar to 1-nanometer precision, resolving molecules only 6 nanometers apart. MINFLUX tracking of single fluorescent proteins increased the temporal resolution and the number of localizations per trace by a factor of 100, as demonstrated with diffusing 30S ribosomal subunits in living Escherichia coli. As conceptual limits have not been reached, we expect this localization modality to break new ground for observing the dynamics, distribution, and structure of macromolecules in living cells and beyond.