使用有机荧光团的定位显微镜中的分子计数
Molecule Counts in Localization Microscopy with Organic Fluorophores.
作者信息
Karathanasis Christos, Fricke Franziska, Hummer Gerhard, Heilemann Mike
机构信息
Institute of Physical and Theoretical Chemistry, Goethe University Frankfurt, Max-von-Laue-Str. 7, 60438, Frankfurt am Main, Germany.
Department of Theoretical Biophysics, Max Planck Institute of Biophysics, Max-von-Laue Str. 3, 60438, Frankfurt am Main, Germany.
出版信息
Chemphyschem. 2017 Apr 19;18(8):942-948. doi: 10.1002/cphc.201601425. Epub 2017 Feb 28.
Single-molecule localization microscopy (SMLM) can be used to count fluorescently labeled molecules even when they are not individually resolved. We demonstrate SMLM molecule counting for nucleic acids labeled with the organic fluorophore Alexa Fluor 647 and imaged under photoswitching conditions. From the observed distributions of the number of fluorophore blinking events, we extract the number of fluorophores per spot using a statistical model. We validate the molecule counting method for single Alexa Fluor 647 fluorophores, and for trimers of Alexa Fluor 647 constructed on a DNA origami structure. This simple counting strategy enables quantitative super-resolution imaging with organic fluorophores.
单分子定位显微镜（SMLM）可用于对荧光标记分子进行计数，即使这些分子无法逐个分辨。我们展示了在光开关条件下对用有机荧光团Alexa Fluor 647标记的核酸进行SMLM分子计数。从观察到的荧光团闪烁事件数量的分布中，我们使用统计模型提取每个斑点的荧光团数量。我们验证了针对单个Alexa Fluor 647荧光团以及在DNA折纸结构上构建的Alexa Fluor 647三聚体的分子计数方法。这种简单的计数策略能够实现有机荧光团的定量超分辨率成像。
Zotero 插件