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Intramolecular Fluorescence Correlation Spectroscopy in a Feedback Tracking Microscope

By Kevin McHale and Hideo Mabuchi

Abstract

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

Topics: Spectroscopy, Imaging, and Other Techniques
Publisher: The Biophysical Society
OAI identifier: oai:pubmedcentral.nih.gov:2895373
Provided by: PubMed Central
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