Article abstract


Nature Photonics 3, 107 - 113 (2008)
Published online: 25 January 2009 | doi:10.1038/nphoton.2008.291

Subject Categories: Imaging and sensing | Biophotonics

Determination of supramolecular structure and spatial distribution of protein complexes in living cells

Valerica Raicu1,2, Michael R. Stoneman1, Russell Fung1, Mike Melnichuk1,4, David B. Jansma1, Luca F. Pisterzi3, Sasmita Rath1, Michael Fox1, James W. Wells3 & Dilano K. Saldin1


Resonant energy transfer from an optically excited donor molecule to a non-excited acceptor molecule residing nearby is widely used to detect molecular interactions in living cells. To date, resonant energy transfer has been used to obtain stoichiometric information, such as the number of proteins forming a complex, for a handful of proteins, but only after performing sequential scans of the emission wavelengths, excitation wavelengths, or sometimes both. During this lengthy process of measurement, the molecular makeup of a cellular region may change, limiting the applicability of resonant energy transfer to the determination of cellular averages. Here, we demonstrate a method for the determination of protein complex size, configuration, and spatial distribution in single living cells. It relies on a spectrally resolved two-photon microscope, a simple but competent theory, and a judicious selection of fluorescent tags. This approach eventually may lead to tracking the dynamics of individual molecular complexes inside living cells.

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  1. Department of Physics, University of Wisconsin-Milwaukee, 1900 E Kenwood Boulevard, Milwaukee, Wisconsin 53211, USA
  2. Department of Biological Sciences, University of Wisconsin-Milwaukee, Milwaukee, Wisconsin 53211, USA
  3. Leslie L. Dan Faculty of Pharmacy, University of Toronto, Toronto, Ontario M5S 3M2, Canada
  4. Present address: Department of Physics, University of Michigan, Ann Arbor, Michigan 48109-1040, USA

Correspondence to: Valerica Raicu1,2 e-mail: vraicu@uwm.edu



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