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arXiv 0908.1177physics.opticsphysics.bio-ph
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反馈跟踪显微镜下的分子内荧光相关光谱

Intramolecular fluorescence correlation spectroscopy in a feedback tracking microscope

Kevin McHale, Hideo Mabuchi

摘要:

我们推导出的反馈跟踪显微镜研究的大分子的形状波动所产生的信号的统计。我们占分子内动态的跟踪系统的响应的影响,并推导出适用于当这些动态是线性的荧光自相关函数的一般表达式。我们发现,跟踪提供了增强的灵敏度,平移扩散,分子大小,异质性和长时间尺度的衰减相比,传统的荧光相关光谱。我们展示了我们的方法,通过使用三维跟踪显微镜研究基因组$λ$ -噬菌体DNA分子与各种荧光标记配置。

英文摘要:

We derive the statistics of the signals generated by shape fluctuations of large molecules studied by feedback tracking microscopy. We account for the influence of intramolecular dynamics on the response of the tracking system, and derive a general expression for the fluorescence autocorrelation function that applies when those dynamics are linear. We show that tracking provides enhanced sensitivity to translational diffusion, molecular size, heterogeneity and long time-scale decays in comparison to traditional fluorescence correlation spectroscopy. We demonstrate our approach by using a three-dimensional tracking microscope to study genomic $λ$-phage DNA molecules with various fluorescence label configurations.

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