Monitoring Membrane Voltage Using Two-Photon Excitation of Fluorescent Voltage-Sensitive Dyes

Functional imaging microscopy based on voltage-sensitive dyes (VSDs) has proven effective for revealing spatiotemporal patterns of activity in vivo and in vitro. Microscopy based on two-photon excitation (TPE) of fluorescent VSDs offers the possibility of three-dimensional recording of membrane potential changes on subcellular length scales hundreds of microns below the brain’s surface. Here we describe progress in monitoring membrane voltage using TPE of VSD fluorescence, and detail an application of this emerging technology in which action potentials (APs) were recorded in single trials from individual mammalian nerve terminals in situ. Prospects for, and limitations of this method are reviewed.

[1]  P. Saggau,et al.  Random-access Multiphoton (ramp) Microscopy Fast Functional Imaging of Single Neurons Using , 2005 .

[2]  W. Webb,et al.  Mechanism of the membrane potential sensitivity of the fluorescent membrane probe merocyanine 540. , 1978, Biochemistry.

[3]  P. Fromherz,et al.  Voltage-sensitive fluorescence of amphiphilic hemicyanine dyes in neuron membrane. , 1993, Biochimica et biophysica acta.

[4]  Walther Akemann,et al.  Engineering and Characterization of an Enhanced Fluorescent Protein Voltage Sensor , 2007, PLoS ONE.

[5]  J Mertz,et al.  Coherent scattering in multi-harmonic light microscopy. , 2001, Biophysical journal.

[6]  L B Cohen,et al.  Optical measurement of membrane potential. , 1978, Reviews of physiology, biochemistry and pharmacology.

[7]  W. Webb,et al.  Multiphoton fluorescence excitation: new spectral windows for biological nonlinear microscopy. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[8]  Jerome Mertz,et al.  Electro-optic response of second-harmonic generation membrane potential sensors , 2003 .

[9]  P. So,et al.  Handbook of Biomedical Nonlinear Optical Microscopy , 2009 .

[10]  C H Wang,et al.  Studies on the mechanism by which cyanine dyes measure membrane potential in red blood cells and phosphatidylcholine vesicles. , 1974, Biochemistry.

[11]  K. Svoboda,et al.  Photon Upmanship: Why Multiphoton Imaging Is More than a Gimmick , 1997, Neuron.

[12]  Bernd Kuhn,et al.  High sensitivity of Stark-shift voltage-sensing dyes by one- or two-photon excitation near the red spectral edge. , 2004, Biophysical journal.

[13]  Vincent A Pieribone,et al.  A genetically targetable fluorescent probe of channel gating with rapid kinetics. , 2002, Biophysical journal.

[14]  L. Cohen,et al.  Optical monitoring of activity from many areas of the in vitro and in vivo salamander olfactory bulb: a new method for studying functional organization in the vertebrate central nervous system , 1983, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[15]  A. Yodh,et al.  One- and two-photon absorption of highly conjugated multiporphyrin systems in the two-photon Soret transition region. , 2009, The Journal of chemical physics.

[16]  W. Douglas.  A Possible Mechanism of Neurosecretion: Release of Vasopressin by Depolarization and its Dependence on Calcium , 1963, Nature.

[17]  Bernd Kuhn,et al.  Anellated hemicyanine dyes in a neuron membrane: Molecular Stark effect and optical voltage recording , 2003 .

[18]  J. A. Fisher.  Linear and non-linear fluorescence imaging of neuronal activity , 2007 .

[19]  B. Salzberg,et al.  Micromolar 4-aminopyridine enhances invasion of a vertebrate neurosecretory terminal arborization: optical recording of action potential propagation using an ultrafast photodiode-MOSFET camera and a photodiode array , 1996, The Journal of general physiology.

[20]  W. Webb,et al.  Nonlinear magic: multiphoton microscopy in the biosciences , 2003, Nature Biotechnology.

[21]  Michinori Ichikawa,et al.  Activity-Dependent Depression of Excitability and Calcium Transients in the Neurohypophysis Suggests a Model of “Stuttering Conduction” , 2003, The Journal of Neuroscience.

[22]  Thomas Knöpfel,et al.  Optical recordings of membrane potential using genetically targeted voltage-sensitive fluorescent proteins. , 2003, Methods.

[23]  A. Zouni,et al.  Voltage sensitivity of the fluorescent probe RH421 in a model membrane system. , 1995, Biophysical Journal.

[24]  Jerome Mertz,et al.  Mechanisms of membrane potential sensing with second-harmonic generation microscopy. , 2003, Journal of biomedical optics.

[25]  Peter T. C. So,et al.  Optical biopsy in high-speed handheld miniaturized multifocal multiphoton microscopy , 2005, SPIE BiOS.

[26]  Leslie M Loew,et al.  The Potential of Dual Camera Systems for Multimodal Imaging of Cardiac Near-infrared Fluorescent Dye Di-4-anbdqbs High-precision Recording of the Action Potential in Isolated Cardiomyocytes Using the Optical Imaging of Voltage and Calcium in Cardiac Cells & Tissues , 2022 .

[27]  Nordmann Jj,et al.  Ultrastructural morphometry of the rat neurohypophysis. , 1977 .

[28]  B. Salzberg,et al.  Optical Recording of Impulses in Individual Neurones of an Invertebrate Central Nervous System , 1973, Nature.

[29]  S. Hell,et al.  Multifocal multiphoton microscopy. , 1998, Optics letters.

[30]  Maria Goeppert-Mayer.  Über Elementarakte mit zwei Quantensprüngen , 1931 .

[31]  Rafael Yuste,et al.  Imaging membrane potential in dendritic spines. , 2006, Proceedings of the National Academy of Sciences of the United States of America.

[32]  Yasushi Okamura,et al.  Phosphoinositide phosphatase activity coupled to an intrinsic voltage sensor , 2005, Nature.

[33]  W. N. Ross,et al.  Optical recording of neuronal activity in an invertebrate central nervous system: simultaneous monitoring of several neurons. , 1977, Journal of neurophysiology.

[34]  B. Salzberg,et al.  Changes in FAD and NADH Fluorescence in Neurosecretory Terminals Are Triggered by Calcium Entry and by ADP Production , 2005, The Journal of Membrane Biology.

[35]  A. Grinvald.  Real-time optical mapping of neuronal activity: from single growth cones to the intact mammalian brain. , 1985, Annual review of neuroscience.

[36]  W. Denk,et al.  Two-photon imaging to a depth of 1000 microm in living brains by use of a Ti:Al2O3 regenerative amplifier. , 2003, Optics letters.

[37]  A. Grinvald,et al.  Spatiotemporal Dynamics of Sensory Responses in Layer 2/3 of Rat Barrel Cortex Measured In Vivo by Voltage-Sensitive Dye Imaging Combined with Whole-Cell Voltage Recordings and Neuron Reconstructions , 2003, The Journal of Neuroscience.

[38]  P. Saggau,et al.  High-speed, random-access fluorescence microscopy: I. High-resolution optical recording with voltage-sensitive dyes and ion indicators. , 1997, Biophysical journal.

[39]  W. Webb,et al.  Two-Photon Fluorescence Excitation Cross Sections of Biomolecular Probes from 690 to 960 nm. , 1998, Applied optics.

[40]  A. Waggoner,et al.  Dye indicators of membrane potential. , 1979, Annual review of biophysics and bioengineering.

[41]  B. Salzberg,et al.  Optical Recording With Single Cell Resolution from a Simple Mammalian Nervous System: Electrical Activity in Ganglia from the Submucous Plexus of the Guinea-Pig Ileum. , 1992, The Biological bulletin.

[42]  B M Salzberg,et al.  Calcium channels that are required for secretion from intact nerve terminals of vertebrates are sensitive to omega-conotoxin and relatively insensitive to dihydropyridines. Optical studies with and without voltage-sensitive dyes , 1989, The Journal of general physiology.

[43]  Gilles Laurent,et al.  A Simple Method to Reconstruct Firing Rates from Dendritic Calcium Signals , 2008, Front. Neurosci..

[44]  C. Peters,et al.  Generation of optical harmonics , 1961 .

[45]  F Bezanilla,et al.  Charge-shift probes of membrane potential. Characterization of aminostyrylpyridinium dyes on the squid giant axon. , 1985, Biophysical journal.

[46]  E. Wolf,et al.  Electromagnetic diffraction in optical systems, II. Structure of the image field in an aplanatic system , 1959, Proceedings of the Royal Society of London. Series A. Mathematical and Physical Sciences.

[47]  Arjun G Yodh,et al.  Two-photon excitation of potentiometric probes enables optical recording of action potentials from mammalian nerve terminals in situ. , 2008, Journal of neurophysiology.

[48]  Rafael Kurtz,et al.  Application of multiline two-photon microscopy to functional in vivo imaging , 2006, Journal of Neuroscience Methods.

[49]  D. Kleinfeld,et al.  Distributed representation of vibrissa movement in the upper layers of somatosensory cortex revealed with voltage‐sensitive dyes , 1996, The Journal of comparative neurology.

[50]  H. Gainer,et al.  Large and rapid changes in light scattering accompany secretion by nerve terminals in the mammalian neurohypophysis , 1985, The Journal of general physiology.

[51]  W. Douglas,et al.  Stimulus—secretion coupling in a neurosecretory organ: the role of calcium in the release of vasopressin from the neurohypophysis , 1964, The Journal of physiology.

[52]  P. Fromherz,et al.  ANNINE-6plus, a voltage-sensitive dye with good solubility, strong membrane binding and high sensitivity , 2007, European Biophysics Journal.

[53]  Leslie M. Loew,et al.  Intracellular long-wavelength voltage-sensitive dyes for studying the dynamics of action potentials in axons and thin dendrites , 2007, Journal of Neuroscience Methods.

[54]  T. Sejnowski,et al.  A Compact Multiphoton 3D Imaging System for Recording Fast Neuronal Activity , 2007, PloS one.

[55]  Yasushi Okamura,et al.  Improving membrane voltage measurements using FRET with new fluorescent proteins , 2008, Nature Methods.

[56]  B. Salzberg,et al.  Novel naphthylstyryl-pyridinium potentiometric dyes offer advantages for neural network analysis , 2004, Journal of Neuroscience Methods.

[57]  B M Salzberg,et al.  Action potentials and frequency-dependent secretion in the mouse neurohypophysis. , 1986, Neuroendocrinology.

[58]  Rafael Yuste,et al.  Imaging neurons : a laboratory manual , 1999 .

[59]  Arjun G Yodh,et al.  In vivo fluorescence microscopy of neuronal activity in three dimensions by use of voltage-sensitive dyes. , 2004, Optics letters.

[60]  Botond Roska,et al.  Tuning FlaSh: redesign of the dynamics, voltage range, and color of the genetically encoded optical sensor of membrane potential. , 2002, Biophysical journal.

[61]  F Bezanilla,et al.  Microsecond response of a voltage-sensitive merocyanine dye: fast voltage-clamp measurements on squid giant axon. , 1993, The Japanese journal of physiology.

[62]  W. Webb,et al.  Measurement of two-photon excitation cross sections of molecular fluorophores with data from 690 to 1050 nm , 1996 .

[63]  D. Senseman,et al.  Optical recording of action potentials from vertebrate nerve terminals using potentiometric probes provides evidence for sodium and calcium components , 1983, Nature.

[64]  P. Marquet,et al.  In vivo local determination of tissue optical properties: applications to human brain. , 1999, Applied optics.

[65]  Leonardo Sacconi,et al.  Optical recording of fast neuronal membrane potential transients in acute mammalian brain slices by second-harmonic generation microscopy. , 2005, Journal of neurophysiology.

[66]  G. Salama,et al.  Properties of New, Long-Wavelength, Voltage-sensitive Dyes in the Heart , 2005, The Journal of Membrane Biology.

[67]  J. White,et al.  Two-photon imaging of spatially extended neuronal network dynamics with high temporal resolution , 2008, Journal of Neuroscience Methods.

[68]  Arjun G. Yodh,et al.  Near infrared two-photon excitation cross-sections of voltage-sensitive dyes , 2005, Journal of Neuroscience Methods.

[69]  W. Denk,et al.  Two-photon laser scanning fluorescence microscopy. , 1990, Science.

[70]  Benjamin Mathieu,et al.  Optical monitoring of neuronal activity at high frame rate with a digital random-access multiphoton (RAMP) microscope , 2008, Journal of Neuroscience Methods.

[71]  C. Sherrington.  Man On His Nature , 1940 .

[72]  D Kleinfeld,et al.  Optical recording of the electrical activity of synaptically interacting Aplysia neurons in culture using potentiometric probes. , 1989, Biophysical journal.

[73]  Peter Saggau,et al.  Compensation of spatial and temporal dispersion for acousto-optic multiphoton laser-scanning microscopy. , 2003, Journal of biomedical optics.

[74]  D. Kleinfeld,et al.  Long-term optical recording of patterns of electrical activity in ensembles of cultured Aplysia neurons. , 1991, Journal of neurophysiology.

[75]  R. Llinás,et al.  The neuronal basis for consciousness. , 1998, Philosophical transactions of the Royal Society of London. Series B, Biological sciences.

[76]  A. Grinvald,et al.  Optical mapping of electrical activity in rat somatosensory and visual cortex , 1985, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[77]  D. Contreras,et al.  Voltage-Sensitive Dye Imaging of Neocortical Spatiotemporal Dynamics to Afferent Activation Frequency , 2001, The Journal of Neuroscience.

[78]  W. N. Ross,et al.  Changes in axon fluorescence during activity: Molecular probes of membrane potential , 1974, The Journal of Membrane Biology.

[79]  Action spectra of electrochromic voltage-sensitive dyes in an intact excitable tissue. , 2008, Journal of biomedical optics.

[80]  R. Llinás.  The intrinsic electrophysiological properties of mammalian neurons: insights into central nervous system function. , 1988, Science.

[81]  D. Kleinfeld,et al.  Functional study of the rat cortical microcircuitry with voltage-sensitive dye imaging of neocortical slices. , 1997, Cerebral cortex.

[82]  B M Salzberg,et al.  Active calcium responses recorded optically from nerve terminals of the frog neurohypophysis , 1985, The Journal of general physiology.

[83]  J. Léger,et al.  Ultrafast random-access scanning in two-photon microscopy using acousto-optic deflectors , 2006, Journal of Neuroscience Methods.

[84]  R. Frostig,et al.  Cortical point-spread function and long-range lateral interactions revealed by real-time optical imaging of macaque monkey primary visual cortex , 1994, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[85]  A. Dunn,et al.  Influence of optical properties on two-photon fluorescence imaging in turbid samples. , 2000, Applied optics.

[86]  T. Z. Teisseyre,et al.  Nonlinear optical potentiometric dyes optimized for imaging with 1064-nm light. , 2007, Journal of biomedical optics.

[87]  T. Knöpfel,et al.  Design and characterization of a DNA‐encoded, voltage‐sensitive fluorescent protein , 2001, The European journal of neuroscience.

[88]  Jerome Mertz,et al.  Two-photon microscopy in brain tissue: parameters influencing the imaging depth , 2001, Journal of Neuroscience Methods.

[89]  T Nielsen,et al.  High efficiency beam splitter for multifocal multiphoton microscopy , 2001, Journal of microscopy.

[90]  Leslie M Loew,et al.  Near-infrared voltage-sensitive fluorescent dyes optimized for optical mapping in blood-perfused myocardium. , 2007, Heart rhythm.

[91]  J. R. Collins.  Change in the Infra-Red Absorption Spectrum of Water with Temperature , 1925 .

[92]  S W Hell,et al.  Two-photon near- and far-field fluorescence microscopy with continuous-wave excitation. , 1998, Optics letters.

[93]  E. Isacoff,et al.  Genetically encoded optical sensors of neuronal activity and cellular function , 2001, Current Opinion in Neurobiology.

[94]  A. Grinvald,et al.  Fluorescence monitoring of electrical responses from small neurons and their processes. , 1983, Biophysical journal.