Palette of fluorinated voltage-sensitive hemicyanine dyes

Year: 2012

Authors: Yan P., Acker C.D., Zhou W.L., Lee P., Bollensdorff C., Negrean A., Lotti J., Sacconi L., Antic S.D., Kohl P., Mansvelder H.D., Pavone F.S., Loew L.M.

Autors Affiliation: R. D. Berlin Center for Cell Analysis and Modeling and Department of Neuroscience, University of Connecticut Health Center, Farmington, CT 06030-6406; Department of Physics, University of Oxford, Oxford OX1 3PU, United Kingdom; National Heart and Lung Institute, Imperial College, London UB9 6JH, United Kingdom; Department of Integrative Neurophysiology, Center for Neurogenomics and Cognitive Research, Neuroscience Campus Amsterdam, Vrije Universiteit Amsterdam, 1081HV, Amsterdam, The Netherlands; European Laboratory for Non-Linear Spectroscopy (LENS) and Department of Physics, University of Florence, 50019 Sesto Fiorentino, Italy; National Institute of Optics, National Research Council (INO-CNR), 50125 Florence, Italy; International Center for Computational Neurophotonics, 50019 Sesto Fiorentino, Italy

Abstract: Optical recording of membrane potential permits spatially resolved measurement of electrical activity in subcellular regions of single cells, which would be inaccessible to electrodes, and imaging of spatiotemporal patterns of action potential propagation in excitable tissues, such as the brain or heart. However, the available voltage-sensitive dyes (VSDs) are not always spectrally compatible with newly available optical technologies for sensing or manipulating the physiological state of a system. Here, we describe a series of 19 fluorinated VSDs based on the hemicyanine class of chromophores. Strategic placement of the fluorine atoms on the chromophores can result in either blue or red shifts in the absorbance and emission spectra. The range of one-photon excitation wavelengths afforded by these new VSDs spans 440-670 nm; the two-photon excitation range is 900-1,340 nm. The emission of each VSD is shifted by at least 100 nm to the red of its one-photon excitation spectrum. The set of VSDs, thus, affords an extended toolkit for optical recording to match a broad range of experimental requirements. We show the sensitivity to voltage and the photostability of the new VSDs in a series of experimental preparations ranging in scale from single dendritic spines to whole heart. Among the advances shown in these applications are simultaneous recording of voltage and calcium in single dendritic spines and optical electrophysiology recordings using two-photon excitation above 1,100 nm.


Volume: 109 (50)      Pages from: 20443  to: 20448

More Information: This work was supported by National Institutes of Health Grant R01 EB001963, the United Kingdom Biotechnology and Biological Sciences Research Council, the British Heart Foundation (BHF), Human Frontier Science Program Grant RGP0027/2009, the Netherlands Research Council Grant NWO 91110019, the European Union Seventh Framework Programme (FP7/2007-2013) under Grant Agreement 284464, the Ente Cassa di Risparmio, Firenze, and the Flagship Nanomax Project. We also thank the University of Oxford Clarendon Fund Scholarship.
KeyWords: fluorescence; microscopy;
DOI: 10.1073/pnas.1214850109

Citations: 135
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