Chemphyschem. 2014 Mar 17;15(4):763-70. doi: 10.1002/cphc.201301080.
The main limiting factor in spatial resolution of localization microscopy is the number of detected photons. Recently we showed that cryogenic measurements improve the photostability of fluorophores, giving access to Angstrom precision in localization of single molecules. Here, we extend this method to colocalize two fluorophores attached to well-defined positions of a double-stranded DNA. By measuring the separations of the fluorophore pairs prepared at different design positions, we verify the feasibility of cryogenic distance measurement with sub-nanometer accuracy. We discuss the important challenges of our method as well as its potential for further improvement and various applications.
定位显微镜空间分辨率的主要限制因素是检测到的光子数量。最近我们表明，低温测量可提高荧光团的光稳定性，从而实现单分子定位的埃精度。在这里，我们将该方法扩展到将两个荧光团附着到双链 DNA 的明确定义位置来进行共定位。通过测量在不同设计位置制备的荧光团对之间的分离，我们验证了低温距离测量具有亚纳米精度的可行性。我们讨论了该方法的重要挑战以及进一步改进和各种应用的潜力。
