欧洲分子生物学实验室Jonas Ries小组取得一项新突破。他们的成果优化了单分子定位显微镜的成像速度和激发光强度。 该项研究成果发表在2020年8月17日出版的《自然—方法学》上。

该团队系统性地量化分析了在单分子定位显微镜中，激发光强度如何影响决定数据质量的两个主要因素——定位精度和标签密度。课题组发现了一个成像速度和质量之间的强的权衡，并展示了为高通量、多色和三维单分子定位显微镜优化的成像方法，该方法能大大提高分辨率和有效的标记效率。系统的评估表明激发光强度对定位显微镜的图像质量有显著影响，并揭示了较低的激发光强度对提高标记效率和定位精度的好处。

研究人员表示，高激光功率是单分子定位显微镜中的常规做法，可以加快数据采集速度。

附：英文原文

Title: Optimizing imaging speed and excitation intensity for single-molecule localization microscopy

Author: Robin Diekmann, Maurice Kahnwald, Andreas Schoenit, Joran Deschamps, Ulf Matti, Jonas Ries

Issue&Volume: 2020-08-17

Abstract: High laser powers are common practice in single-molecule localization microscopy to speed up data acquisition. Here we systematically quantified how excitation intensity influences localization precision and labeling density, the two main factors determining data quality. We found a strong trade-off between imaging speed and quality and present optimized imaging protocols for high-throughput, multicolor and three-dimensional single-molecule localization microscopy with greatly improved resolution and effective labeling efficiency. A systematic evaluation shows that excitation intensity has a dramatic impact on image quality in localization microscopy and reveals the benefits of lower excitation intensity for improved labeling efficiency and localization precision.

DOI: 10.1038/s41592-020-0918-5

Source: https://www.nature.com/articles/s41592-020-0918-5