Single quantum dot tracking based on perceptual grouping using minimal paths in a spatiotemporal volume
Single quantum dot tracking based on perceptual grouping using minimal paths in a spatiotemporal volume
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DOI：
10.1109/tip.2005.852794
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发表时间：
2005-09-01
影响因子：
10.6
通讯作者：
Cohen, LD
中科院分区：
文献类型：
--
作者：
Bonneau, S;Dahan, M;Cohen, LD
半导体量子点 (QD) 是一种新型荧光探针，在超灵敏生物成像方面具有广阔的前景。当在单分子水平上检测时，可以在前所未有的持续时间内观察和追踪活细胞膜中的 QD 标记分子。在荧光图像序列中记录的这些单个分子的运动可以揭示传统整体成像中隐藏的细胞过程动力学的各个方面。然而，由于量子点复杂的光学特性，例如荧光间歇性，这些序列的定量分析具有挑战性并且需要先进的算法。我们在这里提出一种新颖的方法，它不是逐帧分析，而是基于时空体积中的感知分组。通过应用基于图像荧光模型的检测过程，我们首先获得一组非结构化的点。然后，单个分子轨迹被视为从荧光图像堆栈导出的黎曼度量中的最小路径。这些路径是使用快速行进方法的变体计算的，并且需要很少的参数。我们展示了我们的算法在合成数据序列和活神经元膜中单个 QD 标记受体获得的实验测量中跟踪间歇性物体的能力。虽然该方法是为跟踪量子点而开发的，但它可以与任何荧光探针一起使用。
Semiconductor quantum dots (QDs) are new fluorescent probes with great promise for ultrasensitive biological imaging. When detected at the single-molecule level, QD-tagged molecules can be observed and tracked in the membrane of live cells over unprecedented durations. The motion of these individual molecules, recorded in sequences of fluorescence images, can reveal aspects of the dynamics of cellular processes that remain hidden in conventional ensemble imaging. Due to QD complex optical properties, such as fluorescence intermittency, the quantitative analysis of these sequences is, however, challenging and requires advanced algorithms. We present here a novel approach, which, instead of a frame by frame analysis, is based on perceptual grouping in a spatiotemporal volume. By applying a detection process based on an image fluorescence model, we first obtain an unstructured set of points. Individual molecular trajectories are then considered as minimal paths in a Riemannian metric derived from the fluorescence image stack. These paths are computed with a variant of the fast marching method and few parameters are required. We demonstrate the ability of our algorithm to track intermittent objects both in sequences of synthetic data and in experimental measurements obtained with individual QD-tagged receptors in the membrane of live neurons. While developed for tracking QDs, this method can, however, be used with any fluorescent probes.