title: Palette of fluorinated voltage-sensitive hemicyanine dyes.
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.
authors.affiliation: R D Berlin Center for Cell Analysis and Modeling, University of Connecticut Health Center, Farmington, CT 06030-6406, USA.
authors.forename: Ping
authors.initials: P
authors.lastname: Yan
authors.forename: Corey D
authors.initials: CD
authors.lastname: Acker
authors.forename: Wen-Liang
authors.initials: WL
authors.lastname: Zhou
authors.forename: Peter
authors.initials: P
authors.lastname: Lee
authors.forename: Christian
authors.initials: C
authors.lastname: Bollensdorff
authors.forename: Adrian
authors.initials: A
authors.lastname: Negrean
authors.forename: Jacopo
authors.initials: J
authors.lastname: Lotti
authors.forename: Leonardo
authors.initials: L
authors.lastname: Sacconi
authors.forename: Srdjan D
authors.initials: SD
authors.lastname: Antic
authors.forename: Peter
authors.initials: P
authors.lastname: Kohl
authors.forename: Huibert D
authors.initials: HD
authors.lastname: Mansvelder
authors.forename: Francesco S
authors.initials: FS
authors.lastname: Pavone
authors.forename: Leslie M
authors.initials: LM
authors.lastname: Loew
mesh_terms: D000200:Action Potentials; D000818:Animals; D055592:Biophysical Phenomena; D001921:Brain / Q000502:physiology; D020013:Calcium Signaling; D002232:Carbocyanines / Q000737:chemistry*; D004396:Coloring Agents / Q000737:chemistry*; D049229:Dendritic Spines / Q000502:physiology; D005260:Female; D005456:Fluorescent Dyes / Q000737:chemistry; D005461:Fluorine / Q000737:chemistry; D006168:Guinea Pigs; D006321:Heart / Q000502:physiology; D066298:In Vitro Techniques; D051379:Mice; D036641:Microscopy, Fluorescence, Multiphoton / Q000295:instrumentation; D015394:Molecular Structure; D055620:Optical Phenomena; D011689:Purkinje Cells / Q000502:physiology; D051381:Rats; D017208:Rats, Wistar; D013053:Spectrophotometry
chemical_list: D002232:Carbocyanines; D004396:Coloring Agents; D005456:Fluorescent Dyes; D005461:Fluorine