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Bibliographic Details
Main Authors: Raicu, Valerica, Stoneman, Michael R., Fung, Russell, Melnichuk, Mike, Jansma, David B., Pisterzi, Luca, Fox, Michael, Wells, James W., Saldin, Dilano K.
Format: Preprint
Published: 2008
Subjects:
Online Access:https://arxiv.org/abs/0810.2341
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Table of Contents:
  • Resonant Energy Transfer (RET) from an optically excited donor molecule (D) to a non-excited acceptor molecule (A) residing nearby is widely used to detect molecular interactions in living cells. Stoichiometric information, such as the number of proteins forming a complex, has been obtained so far for a handful of proteins, but only after exposing the sample sequentially to at least two different excitation wavelengths. During this lengthy process of measurement, the molecular makeup of a cellular region may change, and this has so far limited the applicability of RET to determination of cellular averages. Here we demonstrate a method for imaging protein complex distribution in living cells with sub-cellular spatial resolution, which relies on a spectrally-resolved two-photon microscope, a simple but competent theory, and a keen selection of fluorescent tags. This technology may eventually lead to tracking dynamics of macromolecular complex formation and dissociation with spatial resolution inside living cells.