Polarization-Encoded Colocalization Microscopy at Cryogenic Temperatures,ACS Photonics

当前位置： X-MOL 学术 › ACS Photonics › 论文详情

Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)

Polarization-Encoded Colocalization Microscopy at Cryogenic Temperatures
ACS Photonics ( IF 6.5 ) Pub Date : 2020-12-22 , DOI: 10.1021/acsphotonics.0c01201
Daniel Böning ₁ , Franz-Ferdinand Wieser _{1,

2} , Vahid Sandoghdar _{1,

2}

Affiliation

Super-resolution localization microscopy is based on determining the positions of individual fluorescent markers in a sample. The major challenge in reaching an ever higher localization precision lies in the limited number of collected photons from single emitters. To tackle this issue, it has been shown that one can exploit the increased photostability at low temperatures, reaching localization precisions in the subnanometer range. Another crucial ingredient of single-molecule super-resolution imaging is the ability to activate an individual emitter within a diffraction-limited spot. Here, we report on the photoblinking behavior of organic dyes at low temperature and elaborate on the limitations of this ubiquitous phenomenon for selecting single molecules. We then show that recording the emission polarization not only provides access to the molecular orientation, but it also facilitates the assignment of photons to individual blinking molecules. Furthermore, we employ periodical modulation of the excitation polarization as a robust method to effectively switch fluorophores. We benchmark each approach by resolving two emitters on different DNA origami structures.

中文翻译：

低温下的偏振编码共定位显微镜

超分辨率定位显微镜基于确定样品中单个荧光标记的位置。达到更高定位精度的主要挑战在于从单个发射器收集的光子数量有限。为了解决这个问题，已经表明人们可以利用在低温下增加的光稳定性，达到亚纳米范围内的定位精度。单分子超分辨率成像的另一个关键要素是能够激活衍射极限点内的单个发射器。在这里，我们报道了有机染料在低温下的光闪烁行为，并详细阐述了这种普遍现象对于选择单分子的局限性。然后，我们表明，记录发射极化不仅可以提供对分子取向的访问，而且还可以促进将光子分配给各个闪烁的分子。此外，我们采用激发极化的周期性调制作为有效切换荧光团的可靠方法。我们通过解析不同DNA折纸结构上的两个发射器来对每种方法进行基准测试。

更新日期：2021-01-20

点击分享查看原文

点击收藏

公开下载

阅读更多本刊新发论文本刊介绍/投稿指南