Feedback controller design for tracking a single fluorescent molecule
Feedback controller design for tracking a single fluorescent molecule
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DOI：
10.1007/s00340-004-1426-5
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发表时间：
2004-03-01
影响因子：
2.1
通讯作者：
Mabuchi, H
中科院分区：
文献类型：
--
作者：
Berglund, AJ;Mabuchi, H
在本文中，我们重新审视了在激光扫描共焦显微镜中跟踪单个荧光分子的问题。我们利用最优控制理论来设计反馈控制器，并使用数值模拟来分析其二维跟踪分子的能力。本文的一个主要主题是包含所有相关的实验限制，包括中等信噪比荧光检测和机电转换阶段的有限带宽响应。这里给出的结果证明了追踪扩散系数高达 0.1 mum(2)/ms 的单个荧光分子的实验可行性。我们表明，对于沿三维适度限制的分子（例如，通过显微镜盖玻片），二维跟踪算法似乎是稳健且有效的。我们期望这项技术能够实现长观察时间和高时间分辨率的单分子实验。
In this paper, we revisit the problem of tracking a single fluorescent molecule in a laser-scanning confocal microscope. We utilize optimal control theory to design a feedback controller and use numerical simulation to analyze its ability to track a molecule in two dimensions. A major theme in this paper is the inclusion of all relevant experimental limitations including moderate signal-to-noise fluorescence detection and the finite-bandwidth response of an electromechanical translation stage. The results presented here demonstrate the experimental feasibility of tracking single fluorescent molecules with diffusion coefficients as large as 0.1 mum(2)/ms. We show that for molecules that are even moderately confined along a third dimension (as, for example, by microscope cover slides), the two-dimensional tracking algorithm appears to be robust and effective. We expect this technology to enable single-molecule experiments with long observation times and high time-resolution.