Tracking Single Particles and Elongated Filaments with Nanometer Precision
Tracking Single Particles and Elongated Filaments with Nanometer Precision
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DOI：
10.1016/j.bpj.2011.04.023
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发表时间：
2011-06-08
影响因子：
3.4
通讯作者：
Diez, Stefan
中科院分区：
文献类型：
--
作者：
Ruhnow, Felix;Zwicker, David;Diez, Stefan
图像处理的最新发展极大地促进了我们对体内和体外生物分子过程的理解。特别是，使用高斯模型来拟合纳米尺寸物体的强度分布，使得它们能够以一纳米范围内的精度进行二维定位。在这里，我们提出了一种算法来精确定位弯曲细丝，其结构的特征是亚分辨直径和微米长度。使用表面固定的微管，用罗丹明荧光标记，当确定细丝中心线时，位置精度接近2 nm，当定位细丝尖端时，位置精度接近9 nm。结合最先进的单粒子跟踪技术，我们将该算法应用于1)电机蛋白质踩在固定微管上，2)解聚微管，3)在电机涂层表面滑动的微管。
Recent developments in image processing have greatly advanced our understanding of biomolecular processes in vitro and in vivo. In particular, using Gaussian models to fit the intensity profiles of nanometer-sized objects have enabled their two-dimensional localization with a precision in the one-nanometer range. Here, we present an algorithm to precisely localize curved filaments whose structures are characterized by subresolution diameters and micrometer lengths. Using surface-immobilized microtubules, fluorescently labeled with rhodamine, we demonstrate positional precisions of similar to 2 nm when determining the filament centerline and similar to 9 nm when localizing the filament tips. Combined with state-of-the-art single particle tracking we apply the algorithm 1), to motor-proteins stepping on immobilized microtubules, 2), to depolymerizing microtubules, and 3), to microtubules gliding over motor-coated surfaces.