Abstract
No methods proposed thus far have the capability to measure molecular flow in live cells at the single molecule level. Here, we review the potentiality of a newly established method based on the spatial correlation of fluorescence fluctuations at a pair of points in the sample (pair correlation method). The pair correlation function (pCF) offers a unique tool to probe the directionality of intracellular traffic, by measuring the accessibility of the cellular landscape and its role in determining the diffusive routes adopted by molecules. The sensitivity of the pCF method toward detection of barriers means that different structural elements of the cell can be tested in terms of penetrability and mechanisms of regulation imparted on molecular flow. This has been recently demonstrated in a series of studies looking at molecular transport inside live cells. Here, we will review the theory behind detection of barriers to molecular flow, the rules to interpret pCF data, and relevant applications to intracellular transport.
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Acknowledgements
The authors are grateful to Enrico Gratton for his invaluable contribution to this work through teaching, continuous motivation and discussion. The authors would also like to thank Michelle Digman for her precious help with planning experiments and discussion of data. This work was supported by NIH-P41-RRO3155, P50-GM076516 and NIH-U54 GM064346 Cell Migration Consortium.
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Hinde, E., Cardarelli, F. Measuring the flow of molecules in cells. Biophys Rev 3, 119–129 (2011). https://doi.org/10.1007/s12551-011-0051-x
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DOI: https://doi.org/10.1007/s12551-011-0051-x