Intramolecular Fluorescence Correlation Spectroscopy in a Feedback Tracking Microscope

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• 作者：Kevin McHale ; Hideo Mabuchi
• 刊名：Biophysical Journal
• 出版年：2010
• 出版时间：7 July 2010
• 年：2010
• 卷：99
• 期：1
• 页码：313-322
• 全文大小：498 K

文摘

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 in comparison to traditional fluorescence correlation spectroscopy, tracking provides enhanced sensitivity to translational diffusion, molecular size, heterogeneity, and long-timescale decays. We demonstrate our approach using a three-dimensional tracking microscope to study genomic λ-phage DNA molecules with various fluorescence label configurations.