Quantification and correction of systematic errors due to detector time-averaging in single-molecule tracking experiments
Quantification and correction of systematic errors due to detector time-averaging in single-molecule tracking experiments
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DOI：
10.1529/biophysj.105.075176
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发表时间：
2006-01-01
影响因子：
3.4
通讯作者：
Salomé, L
中科院分区：
文献类型：
--
作者：
Destainville, N;Salomé, L
单分子跟踪是观察质膜动态组织的一种有效方法。然而，它的使用有一些限制。例如，最近使用数值模拟观察到，探测器曝光时间固有的时间平均效应可能会使微区中分子的表观运动产生偏差。在这里，我们通过分析来解决这个明显受限的问题。我们探讨这一现象，通过计算其对所观察到的扩散系数和域的大小的影响。我们证明了真实的参数可以很容易地从测量的表观参数恢复。有趣的是，我们发现单分子跟踪可以用来探索在小于暴露时间的时间尺度上发生的事件。
Single-molecule tracking is a powerful way to look at the dynamic organization of plasma membranes. However, there are some limitations to its use. For example, it was recently observed, using numerical simulation, that time-averaging effects inherent to the exposure time of detectors are likely to bias the apparent motion of molecules confined in microdomains. Here, we solve this apparently limiting issue analytically. We explore this phenomenon by calculating its effects on the observed diffusion coefficients and domain sizes. We demonstrate that the real parameters can be easily recovered from the measured apparent ones. Interestingly, we find that single-molecule tracking can be used to explore events occurring at a timescale smaller than the exposure time.