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Inferring diffusion in single live cells at the single-molecule level

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JournalPhilosophical Transactions Of The Royal Society Of London Series B - Biological Sciences
DatePublished - 5 Feb 2013
Issue number1611
Volume368
Pages (from-to)1-14
Original languageEnglish

Abstract

The movement of molecules inside living cells is a fundamental feature of biological processes. The ability to both observe and analyse the details of molecular diffusion in vivo at the single-molecule and single-cell level can add significant insight into understanding molecular architectures of diffusing molecules and the nanoscale environment in which the molecules diffuse. The tool of choice for monitoring dynamic molecular localization in live cells is fluorescence microscopy, especially so combining total internal reflection fluorescence with the use of fluorescent protein (FP) reporters in offering exceptional imaging contrast for dynamic processes in the cell membrane under relatively physiological conditions compared with competing single-molecule techniques. There exist several different complex modes of diffusion, and discriminating these from each other is challenging at the molecular level owing to underlying stochastic behaviour. Analysis is traditionally performed using mean square displacements of tracked particles; however, this generally requires more data points than is typical for single FP tracks owing to photophysical instability. Presented here is a novel approach allowing robust Bayesian ranking of diffusion processes to discriminate multiple complex modes probabilistically. It is a computational approach that biologists can use to understand single-molecule features in live cells.

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© 2012 The Authors. This is an author produced version of a paper published in Philosophical Transactions B. Uploaded in accordance with the publisher's self-archiving policy.

    Research areas

  • Cell Tracking, Cells, DNA, Viral, Diffusion, Fluorescence, Fluorescent Dyes, HIV-1, Metal Nanoparticles, Models, Biological, Nanotechnology, Reproducibility of Results, Spectrum Analysis, Raman

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