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

Robson, Alex, Burrage, Kevin, & Leake, Mark C. (2012) Inferring diffusion in single live cells at the single-molecule level. Philosophical Transactions of the Royal Society B: Biological Sciences, 368(1611), p. 20120029.

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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 diffus- ing 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 mem- brane 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 mol- ecular 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 dis-criminate 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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10 citations in Scopus
28 citations in Web of Science®
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ID Code: 65851
Item Type: Journal Article
Refereed: Yes
Keywords: Diffusion, Confinement, Fluorescent proteins, In vivo imaging, Single particle tracking, Membrane heterogeneity
DOI: 10.1098/rstb.2012.0029
ISSN: 1471-2970
Divisions: Current > Schools > School of Mathematical Sciences
Current > QUT Faculties and Divisions > Science & Engineering Faculty
Deposited On: 09 Jan 2014 04:28
Last Modified: 09 Jan 2014 04:29

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