floating image of dye vials

Research Application Overview

Voltage-sensitive dyes (VSDs) are small molecules that convert changes in voltage inside cells to visible changes in fluorescence. Voltage-sensitive dye imaging is used to record these changes in fluorescence, usually with a microscope.

They are an important tool for researchers who need to measure electrical signaling in cells for several main purposes:

Neuroscience Research

Neuroscience researchers study neurological disorders such as Alzheimer’s and Autism

Microscopic view of fluorescent neural activity with an overlaid graph showing activity spike; bright areas indicate active neurons.

Commonly Used:

Cardiac Research

Cardiac researchers study conditions such as Arrhythmogenic Cardiomyopathy

Commonly Used:

Fluorescent image of cardiac tissue from Peter Lee at Essel R&D. Single-sensor system for spatially resolved, continuous, and multiparametric optical mapping of cardiac tissue.

Image Courtesy of Peter Lee, Essel R&D

High Throughput Screening

Drug developers test the efficacy or safety of new therapeutic compounds

Graphical representation of different cardiac rhythms in human iPSC-CMs, showcasing normal beating, and Type A, B, C, and T arrhythmias, using Di-4-ANEPPS dye for visualization.

Commonly Used:

Blinova, Ksenia et al. (2017) Pubmed

New & Novel Applications

Researchers use cancer cell lines to track hyperpolarization propagation

Microscopic image showing cells with numbered regions, with corresponding line graphs representing data analysis of each region. Voltage imaging reveals the dynamic and propagating electrical signatures of human breast cancer cells.

Commonly Used:

 Related Literature:

  • 1. Herron, T. J., P. Lee, and J. Jalife. 2012. Optical imaging of voltage and calcium in cardiac cells & tissues. Circ Res. Pubmed

    2. Peterka, D. S., H. Takahashi, and R. Yuste. 2011. Imaging voltage in neurons. Neuron. Pubmed

    3. Loew, L. M. 2015. Design and Use of Organic Voltage Sensitive Dyes. Adv Exp Med Biol. Pubmed

  • 1. Crocini, C., R. Coppini, C. Ferrantini, P. Yan, L. M. Loew, C. Tesi, E. Cerbai, C. Poggesi, F. S. Pavone, and L. Sacconi. 2014. Defects in T-tubular electrical activity underlie local alterations of calcium release in heart failure. Proc Natl Acad Sci U S A 111(42):15196-15201. Pubmed

    2. Lee, P., C. Bollensdorff, T. A. Quinn, J. P. Wuskell, L. M. Loew, and P. Kohl. 2011. Single-sensor system for spatially resolved, continuous, and multiparametric optical mapping of cardiac tissue. Heart Rhythm 8(9):1482-1491. Pubmed

  • 1. Acker, C. D., E. Hoyos, and L. M. Loew. 2016. EPSPs Measured in Proximal Dendritic Spines of Cortical Pyramidal Neurons. eNeuro 3(2). Pubmed

    2. Rowan, M. J., E. Tranquil, and J. M. Christie. 2014. Distinct kv channel subtypes contribute to differences in spike signaling properties in the axon initial segment and presynaptic boutons of cerebellar interneurons. J Neurosci 34(19):6611-6623. Pubmed

  • 1. Hortigon-Vinagre, M. P., V. Zamora, F. L. Burton, J. Green, G. A. Gintant, and G. L. Smith. 2016. The Use of Ratiometric Fluorescence Measurements of the Voltage Sensitive Dye Di-4-ANEPPS to Examine Action Potential Characteristics and Drug Effects on Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes. Toxicol Sci 154(2):320-331. Pubmed

    2. Blinova, K., J. Stohlman, J. Vicente, D. Chan, L. Johannesen, M. P. Hortigon-Vinagre, V. Zamora, G. Smith, W. J. Crumb, L. Pang, B. Lyn-Cook, J. Ross, M. Brock, S. Chvatal, D. Millard, L. Galeotti, N. Stockbridge, and D. G. Strauss. 2017. Comprehensive Translational Assessment of Human-Induced Pluripotent Stem Cell Derived Cardiomyocytes for Evaluating Drug-Induced Arrhythmias. Toxicol Sci 155(1):234-247. Pubmed

  • 1. Acker, C. D., E. Hoyos, and L. M. Loew. 2016. EPSPs Measured in Proximal Dendritic Spines of Cortical Pyramidal Neurons. eNeuro 3(2). Pubmed

    2. Crocini, C., R. Coppini, C. Ferrantini, P. Yan, L. M. Loew, C. Tesi, E. Cerbai, C. Poggesi, F. S. Pavone, and L. Sacconi. 2014. Defects in T-tubular electrical activity underlie local alterations of calcium release in heart failure. Proc Natl Acad Sci U S A 111(42):15196-15201. Pubmed

    3. Yan, P., C. D. Acker, W. L. Zhou, P. Lee, C. Bollensdorff, A. Negrean, J. Lotti, L. Sacconi, S. D. Antic, P. Kohl, H. D. Mansvelder, F. S. Pavone, and L. M. Loew. 2012. Palette of fluorinated voltage-sensitive hemicyanine dyes. Proceedings of the National Academy of Sciences of the United States of America 109(50):20443-20448. Pubmed

    4. Rowan, M. J., E. Tranquil, and J. M. Christie. 2014. Distinct kv channel subtypes contribute to differences in spike signaling properties in the axon initial segment and presynaptic boutons of cerebellar interneurons. J Neurosci 34(19):6611-6623. Pubmed

  • 1. Zhou, W. L., P. Yan, J. P. Wuskell, L. M. Loew, and S. D. Antic. 2007. Intracellular long-wavelength voltage-sensitive dyes for studying the dynamics of action potentials in axons and thin dendrites. J Neurosci Methods 164(2):225-239. Pubmed

    2. Warren, M., K. W. Spitzer, B. W. Steadman, T. D. Rees, P. Venable, T. Taylor, J. Shibayama, P. Yan, J. P. Wuskell, L. M. Loew, and A. V. Zaitsev. 2010. High-precision recording of the action potential in isolated cardiomyocytes using the near-infrared fluorescent dye di-4-ANBDQBS. Am J Physiol Heart Circ Physiol 299(4):H1271-1281. Pubmed

    3. Matiukas, A., B. G. Mitrea, M. Qin, A. M. Pertsov, A. G. Shvedko, M. D. Warren, A. V. Zaitsev, J. P. Wuskell, M. D. Wei, J. Watras, and L. M. Loew. 2007. Near-infrared voltage-sensitive fluorescent dyes optimized for optical mapping in blood-perfused myocardium. Heart Rhythm 4(11):1441-1451. Pubmed

    4. Lee, P., C. Bollensdorff, T. A. Quinn, J. P. Wuskell, L. M. Loew, and P. Kohl. 2011. Single-sensor system for spatially resolved, continuous, and multiparametric optical mapping of cardiac tissue. Heart Rhythm 8(9):1482-1491. Pubmed

  • 1. Bachtel, A. D., R. A. Gray, J. M. Stohlman, E. B. Bourgeois, A. E. Pollard, and J. M. Rogers. 2011. A novel approach to dual excitation ratiometric optical mapping of cardiac action potentials with di-4-ANEPPS using pulsed LED excitation. IEEE Trans Biomed Eng 58(7):2120-2126. Pubmed

    2. Fluhler, E., V. G. Burnham, and L. M. Loew. 1985. Spectra, membrane binding, and potentiometric responses of new charge shift probes. Biochemistry 24(21):5749-5755. Pubmed

    3. Montana, V., D. L. Farkas, and L. M. Loew. 1989. Dual-wavelength ratiometric fluorescence measurements of membrane potential. Biochemistry 28(11):4536-4539. Pubmed

    4. Loew, L. M., L. B. Cohen, J. Dix, E. N. Fluhler, V. Montana, G. Salama, and J. Y. Wu. 1992. A naphthyl analog of the aminostyryl pyridinium class of potentiometric membrane dyes shows consistent sensitivity in a variety of tissue, cell, and model membrane preparations. J Membr Biol 130(1):1-10. Pubmed