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A multi-emitter fitting algorithm for potential live cell super-resolution imaging over a wide range of molecular densities
Journal of Microscopy ( IF 2 ) Pub Date : 2018-05-25 , DOI: 10.1111/jmi.12714
T Takeshima 1 , T Takahashi 1 , J Yamashita 1 , Y Okada 2, 3 , S Watanabe 1
Affiliation  

Multi‐emitter fitting algorithms have been developed to improve the temporal resolution of single‐molecule switching nanoscopy, but the molecular density range they can analyse is narrow and the computation required is intensive, significantly limiting their practical application. Here, we propose a computationally fast method, wedged template matching (WTM), an algorithm that uses a template matching technique to localise molecules at any overlapping molecular density from sparse to ultrahigh density with subdiffraction resolution. WTM achieves the localization of overlapping molecules at densities up to 600 molecules μm–2 with a high detection sensitivity and fast computational speed. WTM also shows localization precision comparable with that of DAOSTORM (an algorithm for high‐density super‐resolution microscopy), at densities up to 20 molecules μm–2, and better than DAOSTORM at higher molecular densities. The application of WTM to a high‐density biological sample image demonstrated that it resolved protein dynamics from live cell images with subdiffraction resolution and a temporal resolution of several hundred milliseconds or less through a significant reduction in the number of camera images required for a high‐density reconstruction. WTM algorithm is a computationally fast, multi‐emitter fitting algorithm that can analyse over a wide range of molecular densities. The algorithm is available through the website. https://doi.org/10.17632/bf3z6xpn5j.1

中文翻译:

一种用于在广泛分子密度范围内进行潜在活细胞超分辨率成像的多发射器拟合算法

已经开发了多发射器拟合算法来提高单分子开关纳米显微镜的时间分辨率,但它们可以分析的分子密度范围很窄,所需的计算量很大,极大地限制了它们的实际应用。在这里,我们提出了一种计算速度快的方法,楔形模板匹配 (WTM),该算法使用模板匹配技术将分子定位在任何重叠分子密度,从稀疏到超高密度,具有亚衍射分辨率。WTM 实现了密度高达 600 个分子 μm-2 的重叠分子的定位,具有高检测灵敏度和快速计算速度。WTM 还显示出与 DAOSTORM(一种用于高密度超分辨率显微镜的算法)相当的定位精度,密度高达 20 个分子 μm-2,并且在更高的分子密度下优于 DAOSTORM。WTM 在高密度生物样本图像中的应用表明,它通过显着减少高密度生物样本图像所需的相机图像数量,从具有亚衍射分辨率和数百毫秒或更短时间分辨率的活细胞图像中解析蛋白质动力学。密度重建。WTM 算法是一种计算速度快的多发射器拟合算法,可以在很宽的分子密度范围内进行分析。该算法可通过网站获得。https://doi.org/10.17632/bf3z6xpn5j.1 WTM 在高密度生物样本图像中的应用表明,它通过显着减少高密度生物样本图像所需的相机图像数量,从具有亚衍射分辨率和数百毫秒或更短时间分辨率的活细胞图像中解析蛋白质动力学。密度重建。WTM 算法是一种计算速度快的多发射器拟合算法,可以在很宽的分子密度范围内进行分析。该算法可通过网站获得。https://doi.org/10.17632/bf3z6xpn5j.1 WTM 在高密度生物样本图像中的应用表明,它通过显着减少高密度生物样本图像所需的相机图像数量,从具有亚衍射分辨率和数百毫秒或更短时间分辨率的活细胞图像中解析蛋白质动力学。密度重建。WTM 算法是一种计算速度快的多发射器拟合算法,可以在很宽的分子密度范围内进行分析。该算法可通过网站获得。https://doi.org/10.17632/bf3z6xpn5j.1 多发射器拟合算法,可以在很宽的分子密度范围内进行分析。该算法可通过网站获得。https://doi.org/10.17632/bf3z6xpn5j.1 多发射器拟合算法,可以在很宽的分子密度范围内进行分析。该算法可通过网站获得。https://doi.org/10.17632/bf3z6xpn5j.1
更新日期:2018-05-25
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