Fluorescence resonance energy transfer (FRET) is widely recognized as being the main fluorescence tool to obtain quantitative information about the dynamics and structure of macromolecules. Yet, because FRET depends on both the distance (dDA) between a pair of donor and acceptor and the internal dynamics of the macromolecules that control the displacement of the dyes, it is rarely applied to macromolecules labeled with more than a single energy donor and acceptor separated by a single contour length. Indeed, beside end-labeled linear oligomers, FRET is hardly ever used for quantitative studies on large macromolecules with more complex architectures due to intractable mathematical complications associated with the difficulty of tracking an a priori unknown dDA distribution that varies as a function of time. This presentation will demonstrate that these mathematical complications vanish when dealing with pyrene excimer formation/fluorescence (PEF) since PEF occurs on contact which removes the FRET requirement of dealing with a time-dependent dDA distribution. The effectiveness of PEF for the characterization of pyrene-labeled macromolecules will be illustrated with a series of pyrene-labeled dendrimers and a series of pyrene-labeled polymeric bottle brushes.

Pyrene excimer fluorescence yields the same structural and dynamic information on macromolecules as FRET but in a mathematically much simpler manner

256th National Meeting (2018)

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