New statistical tools for single molecule experiments

描述（由申请人提供）：单分子（SM）测量在世界各地的研究实验室中正迅速变得司空见惯，并对许多研究领域做出贡献，因为它们能够提供对以前由于整体测量中存在的系综平均而难以处理的现象的洞察。特别是构象异构系统的动力学正受益于单分子研究。蛋白质折叠和构象动力学、酶学、核酶功能、细菌光捕获和蛋白质-核酸相互作用只是从SM技术的应用中受益的复杂系统的几个例子。然而，由于缺乏统一的数据分析和解释，SM结果的影响有所减轻。拟议的研究重点是SM荧光测量以及如何将实验设计，分析和期望置于坚实的统计和理论基础上。提出了三个具体目标：1。利用信息论确定SM实验的基本极限。2.开发基于隐马尔可夫模型的严格统计分析方法。 3.实现方法作为面向用户的通用数据分析包的补充。 这项研究对健康的重要性在于它对许多正在进行的SM生物系统研究的贡献。SM测量正在彻底改变我们对化学生物学中许多问题的方法，但它们仍然是基于仅对批量样品的集合测量有效的假设来解释和设计的。这可能导致收集的数据无法使用传统方法进行充分解释。一个一致的SM测量理论框架将是该领域的重要一步。 目标1将提供一个理论框架，可用于实验设计，因为它提供了测量的能力，使系统的属性推断的限制。它还将提供基准（Cramr-Rao界）来判断数据约简方法。目标2开发算法和核心代码，以实现统计上严格的数据分析方法，允许无偏估计系统参数，精度接近Cramr-Rao界包括意义不确定性估计。目标3为实验设计和分析提供了可用的工具，允许其他研究人员利用这些方法进行自己的研究。

DESCRIPTION (provided by applicant): Single molecule (SM) measurements are rapidly becoming commonplace in research laboratories around the world and are contributing to many areas of investigation because of their ability to provide insight into phenomena that were previously intractable because of the ensemble averaging present in bulk measurements. In particular the dynamics of conformationally heterogeneous systems are benefiting from single-molecule studies. Protein folding and conformational dynamics, enzymology, ribozyme function, bacterial light harvesting, and protein-nucleic acid interactions are just a few examples of complex systems that have benefited from the application of SM techniques. However, the impact of SM results has been mitigated by the lack of uniform data analysis and interpretation. The proposed research focuses on SM fluorescence measurements and how to place the experimental design, analysis, and expectations onto solid statistical and theoretical ground. Three specific aims are proposed: 1. Use information theory to determine the fundamental limits of SM experiments. 2. Develop statistically rigorous analysis methods based on hidden Markov models. 3. Implement methods as user-oriented additions to common data analysis packages. The significance to health of this research is through its contribution to the many ongoing SM investigations into biological systems. SM measurements are revolutionizing our approach to many problems in chemical biology, yet they are still being interpreted and designed based on assumptions that are only valid for ensemble measurements of bulk samples. This can result in collection of data that cannot be adequately interpreted using traditional methods. A consistent theoretical framework for SM measurements would be a significant step forward for the field. Aim 1 will provide a theoretical framework that can be used for experimental design as it provides the limit of the measurement's ability to make inferences about the properties of the system. It will also provide the benchmark (the Cramr-Rao bound) by which to judge data reduction methods. Aim 2 develops the algorithms and core codes to implement statistically rigorous methods of data analysis that allow unbiased estimation of system parameters with accuracy approaching the Cramr-Rao bound including meaning uncertainty estimates. Aim 3 provides useable tools for experimental design and analysis to allow other investigators to exploit these methods for their own research.