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Synthesis and use of an in-solution ratiometric fluorescent viscosity sensor

Nature Protocols volume 2, pages 227–236 (2007)Cite this article

Abstract

A procedure for the synthesis of a ratiometric viscosity fluorescent sensor is described in this protocol. The essential requirement for the design of this sensor is the attachment of a primary fluorophore that has both a viscosity-independent fluorescence emission (coumarin dye shown in blue) and an emission from a fluorophore that exhibits viscosity-dependent fluorescent quantum yield (p-amino cinnamonitrile dye shown in red). The use of sensor 1 in viscosity measurements involves solubilization in a liquid of interest and excitation of the primary fluorophore at λex = 360 nm. The secondary fluorophore is simultaneously excited via resonance energy transfer. The ratio of the fluorescent emission of the secondary over the primary fluorophore provides a fast and precise measurement of the viscosity of the solvent. The synthesis of compound 1 using commercially available materials can be completed within 5 d.

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Figure 1: Synthesis of sensor 1.
Figure 2: Typical spectra and typical behavior of emission maxima in fluids of different viscosity (mixtures of ethylene glycol and glycerol).

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Acknowledgements

Financial support by the NIH (1R33 018399) and the UCSD is gratefully acknowledged.

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Authors and Affiliations

  1. Department of Chemistry and Biochemistry, University of California, San Diego, La Jolla, 92093-0358, California, USA

    Derek Fischer & Emmanuel A Theodorakis

  2. Department of Biological Engineering, University of Missouri-Columbia, Columbia, 65211, Missouri, USA

    Mark A Haidekker

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  1. Derek Fischer
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  2. Emmanuel A Theodorakis
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  3. Mark A Haidekker
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Correspondence to Derek Fischer or Mark A Haidekker.

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Fischer, D., Theodorakis, E. & Haidekker, M. Synthesis and use of an in-solution ratiometric fluorescent viscosity sensor. Nat Protoc 2, 227–236 (2007). https://doi.org/10.1038/nprot.2006.455

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