Xu Xinzhu, Jia Shu, Xi Peng
Department of Biomedical Engineering, College of Future Technology, Peking University, Beijing, 100871, China.
Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University, Atlanta, Georgia, 30332, USA.
Light Sci Appl. 2022 Oct 10;11(1):293. doi: 10.1038/s41377-022-00983-6.
A donut excitation moves around a single molecule with a zigzag configuration lattice by lattice. Such a method implemented in scanning fluorescence microscopy simplifies the conventional MINFLUX process. Consisting of hollow zero-intensity excitation, single-pixel detection, time-correlated single photon counting, and drift stabilization, the system achieves localization precision and resolution very close to conventional MINFLUX theoretically and experimentally. An averaged high-SNR reference, and pixel-registered intensity from a single molecule is essential to reconstruct localization in maximum likelihood estimation. With performance reaching nearly conventional MINFLUX's, the proposed raster-scanning MINFLUX can inspire researchers expertized in STED or confocal setup to quickly transform to MINFLUX and develop for further exploring on bio-specimens or optical applications.
甜甜圈形激发以锯齿形配置逐格围绕单个分子移动。这种在扫描荧光显微镜中实现的方法简化了传统的MINFLUX过程。该系统由中空零强度激发、单像素检测、时间相关单光子计数和漂移稳定组成，在理论和实验上都实现了非常接近传统MINFLUX的定位精度和分辨率。在最大似然估计中，平均高信噪比参考以及来自单个分子的像素配准强度对于重建定位至关重要。所提出的光栅扫描MINFLUX性能接近传统MINFLUX，能够激励精通STED或共聚焦设置的研究人员快速转向MINFLUX，并进一步开发用于生物样本或光学应用的探索。
