Applied Physics-Biophysics and Center for NanoScience, Ludwig-Maximilians-Universität, Amalienstrasse 54, D-80799 München, Germany.
Nano Lett. 2010 Feb 10;10(2):645-51. doi: 10.1021/nl903730r.
In this paper we experimentally combine a recently developed AFM-based molecule-by-molecule assembly (single-molecule cut-and-paste, SMCP) with subdiffraction resolution fluorescence imaging. Using "Blink-Microscopy", which exploits the fluctuating emission of single molecules for the reconstruction of superresolution images, we resolved SMCP assembled structures with features below the diffraction limit. Artificial line patterns then served as calibration structures to characterize parameters, such as the labeling density, that can influence resolution of Blink-Microscopy besides the localization precision of a single molecule. Finally, we experimentally utilized the adjustability of blink parameters to demonstrate the general connection of photophysical parameters with spatial resolution and acquisition time in superresolution microscopy.
在本文中，我们实验性地将最近开发的基于原子力显微镜的分子级组装（单分子切割和粘贴，SMCP）与亚衍射分辨率荧光成像相结合。利用“闪烁显微镜”（Blink-Microscopy），它利用单个分子的波动发射来重建超分辨率图像，我们解析了具有低于衍射极限的特征的 SMCP 组装结构。然后，人工线图案作为校准结构来表征可以影响 Blink-Microscopy 分辨率的参数，除了单个分子的定位精度之外，这些参数包括标记密度。最后，我们实验性地利用闪烁参数的可调节性来证明光物理参数与超分辨率显微镜中的空间分辨率和采集时间之间的一般联系。
