Skip to main content
SpringerLink
Log in

Second Harmonic Imaging of Membrane Potential

  • Chapter
Membrane Potential Imaging in the Nervous System and Heart

Part of the book series: Advances in Experimental Medicine and Biology ((AEMB,volume 859))

Abstract

The non-linear optical effect known as second harmonic generation (SHG) has been recognized since the earliest days of the laser. But it has only been in the last 20 years that it has begun to emerge as a viable microscope imaging contrast mechanism for visualization of cell and tissue structure and function. This is because only small modifications are required to equip a standard laser scanning 2-photon microscope for second harmonic imaging microscopy (SHIM). SHG signals from certain membrane-bound dyes are highly sensitive to membrane potential, indicating that SHIM may become a valuable probe of cell physiology. However, for the current generation of dyes and microscopes, the small signal size limits the number of photons that can be collected during the course of a fast action potential. Better dyes and optimized microscope optics could ultimately lead to the ability to image neuronal electrical activity with SHIM.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
Chapter
USD 29.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 129.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 169.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 169.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  • Araya R, Jiang J, Eisenthal KB, Yuste R (2006) The spine neck filters membrane potentials. Proc Natl Acad Sci U S A 103:17961–17966

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Araya R, Nikolenko V, Eisenthal KB, Yuste R (2007) Sodium channels amplify spine potentials. Proc Natl Acad Sci U S A 104:12347–12352

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Ben-Oren I, Peleg G, Lewis A, Minke B, Loew LM (1996) Infrared nonlinear optical measurements of membrane potential in photoreceptor cells. Biophys J 71:1616–1620

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Bianchini P, Diaspro A (2008) Three-dimensional (3D) backward and forward second harmonic generation (SHG) microscopy of biological tissues. J Biophotonics 1:443–450

    Article  PubMed  Google Scholar 

  • Bouevitch O, Lewis A, Pinevsky I, Wuskell JP, Loew LM (1993) Probing membrane potential with non-linear optics. Biophys J 65:672–679

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Campagnola P, Loew LM (2003) Second-harmonic imaging microscopy for visualizing biomolecular arrays in cells, tissues and organisms. Nat Biotechnol 21:1356–1360

    Article  CAS  PubMed  Google Scholar 

  • Campagnola PJ, Clark HA, Mohler WA, Lewis A, Loew LM (2001) Second harmonic imaging microscopy of living cells. J Biomed Opt 6:277–286

    Article  CAS  PubMed  Google Scholar 

  • Campagnola PJ, Wei M-d, Lewis A, Loew LM (1999) High resolution optical imaging of live cells by second harmonic generation. Biophys J 77:3341–3349

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Clark HA, Campagnola PJ, Wuskell JP, Lewis A, Loew LM (2000) Second harmonic generation properties of fluorescent polymer encapsulated gold nanoparticles. J Am Chem Soc 122:10234–10235

    Article  CAS  Google Scholar 

  • Cole JM, Kreiling S (2002) Exploiting structure/property relationships in organic non-linear optical materials: developing strategies to realize the potential of TCNQ derivatives. CrystEngComm 4:232–238

    Article  CAS  Google Scholar 

  • Conboy JC, Richmond GL (1997) Examination of the electrochemical interface between two immiscible electrolye solutions by second harmonic generation. J Phys Chem B 101:983–990

    Article  CAS  Google Scholar 

  • Denk W, Strickler JH, Webb WW (1990) Two-photon laser scanning fluorescence microscopy. Science 248:73–76

    Article  CAS  PubMed  Google Scholar 

  • Dirk CW, Twieg RJ, Wagniere J (1986) The Contribution of Pi Electrons to Second Harmonic Generation in Organic Molecule. J Am Chem Soc 108:5387–5395

    Article  CAS  Google Scholar 

  • Dombeck DA, Blanchard-Desce M, Webb WW (2004) Optical recording of action potentials with second-harmonic generation microscopy. J Neurosci 24:999–1003

    Article  CAS  PubMed  Google Scholar 

  • Dombeck DA, Sacconi L, Blanchard-Desce M, Webb WW (2005) Optical recording of fast neuronal membrane potential transients in acute mammalian brain slices by second-harmonic generation microscopy. J Neurophysiol 94:3628–3636

    Article  PubMed  Google Scholar 

  • Fluhler E, Burnham VG, Loew LM (1985) Spectra, membrane binding and potentiometric responses of new charge shift probes. Biochemistry 24:5749–5755

    Article  CAS  PubMed  Google Scholar 

  • Freund I, Deutsch M, Sprecher A (1986) Connective tissue polarity. Optical second-harmonic microscopy, crossed-beam summation, and small-angle scattering in rat-tail tendon. Biophys J 50:693–712

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Huang J, Lewis A (1989) Determination of the absolute orientation of the retinyldiene chromophore in purple membrane by a second harmonic interference technique. Biophys J 55:835–842

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Huang JY, Lewis A, Loew LM (1988) Non-linear optical properties of potential sensitive styryl dyes. Biophys J 53:665–670

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Jiang J, Eisenthal KB, Yuste R (2007) Second harmonic generation in neurons: electro-optic mechanism of membrane potential sensitivity. Biophys J 93:L26–L28

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Millard AC, Campagnola PJ, Mohler W, Lewis A, Loew LM (2003a) Second harmonic imaging microscopy. In: Marriott G, Parker I (eds) Methods in enzymology, vol. 361B. Academic, San Diego, pp 47–69

    Google Scholar 

  • Millard AC, Jin L, Lewis A, Loew LM (2003b) Direct measurement of the voltage sensitivity of second-harmonic generation from a membrane dye in patch-clamped cells. Opt Lett 28:1221–1223

    Article  CAS  PubMed  Google Scholar 

  • Millard AC, Jin L, Wei M-d, Wuskell JP, Lewis A, Loew LM (2004) Sensitivity of second harmonic generation from styryl dyes to trans-membrane potential. Biophys J 86:1169–1176

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Millard AC, Jin L, Wuskell JP, Boudreau DM, Lewis A, Loew LM (2005a) Wavelength- and time-dependence of potentiometric non-linear optical signals from styryl dyes. J Membr Biol 208:103–111

    Article  CAS  PubMed  Google Scholar 

  • Millard AC, Lewis A, Loew LM (2005b) Second harmonic imaging of membrane potential. In: Yuste R, Konnerth A (eds) Imaging in neuroscience and development. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, pp 463–474

    Google Scholar 

  • Mohler W, Millard AC, Campagnola PJ (2003) Second harmonic generation imaging of endogenous structural proteins. Methods 29:97–109

    Article  CAS  PubMed  Google Scholar 

  • Moreaux L, Pons T, Dambrin V, Blanchard-Desce M, Mertz J (2003) Electro-optic response of second-harmonic generation membrane potential sensors. Opt Lett 28:625–627

    Article  CAS  PubMed  Google Scholar 

  • Moreaux L, Sandre O, Blanchard-desce M, Mertz J (2000a) Membrane imaging by simultaneous second-harmonic generation and two-photo microscopy. Opt Lett 25:320–322

    Article  CAS  PubMed  Google Scholar 

  • Moreaux L, Sandre O, Mertz J (2000b) Membrane imaging by second harmonic generation microscopy. J Opt Soc Am B 17:1685–1694

    Article  CAS  Google Scholar 

  • Morley JO (1988) Non-linear optical properties of organic molecules. 7. Calculated hyperpolarizabilities of azulenes and sesquifulvalene. J Am Chem Soc 110:7660–7663

    Article  CAS  Google Scholar 

  • Nemet BA, Nikolenko V, Yuste R (2004) Second harmonic imaging of membrane potential of neurons with retinal. J Biomed Opt 9:873–881

    Article  PubMed  Google Scholar 

  • Nicoud JF, Twieg RJ (1987) Design and synthesis of organic molecular compounds for efficient second-harmonic generation. In: Chemla DS, Zyss J (eds) Nonlinear optical properties of organic molecules and crystals, vol. 1. Academic, Orlando, pp 227–296

    Chapter  Google Scholar 

  • Nuriya M, Jiang J, Nemet B, Eisenthal KB, Yuste R (2006) Imaging membrane potential in dendritic spines. Proc Natl Acad Sci U S A 103:786–790

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Nuriya M, Yasui M (2010) Membrane potential dynamics of axons in cultured hippocampal neurons probed by second-harmonic-generation imaging. J Biomed Opt 15:020503

    Article  PubMed  Google Scholar 

  • Pons T, Moreaux L, Mongin O, Blanchard-Desce M, Mertz J (2003) Mechanisms of membrane potential sensing with second-harmonic generation microscopy. J Biomed Opt 8:428–431

    Article  CAS  PubMed  Google Scholar 

  • Rama S, Vetrivel L, Semyanov A (2010) Second-harmonic generation voltage imaging at subcellular resolution in rat hippocampal slices. J Biophotonics 3:784–790

    Article  PubMed  Google Scholar 

  • Sacconi L, Dombeck DA, Webb WW (2006) Overcoming photodamage in second-harmonic generation microscopy: real-time optical recording of neuronal action potentials. Proc Natl Acad Sci U S A 103:3124–3129

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Sheppard C, Kompfner R, Gannaway J, Walsh D (1977) IEEE J Quant Electron 13D:100E

    Google Scholar 

  • Teisseyre TZ, Millard AC, Yan P, Wuskell JP, Wei M-d, Lewis A, Loew LM (2007) Non-linear optical potentiometric dyes optimized for imaging with 1064 nm light. J Biomed Opt 12:044001–044008

    Article  PubMed  Google Scholar 

  • Theer P, Denk W, Sheves M, Lewis A, Detwiler PB (2011) Second-harmonic generation imaging of membrane potential with retinal analogues. Biophys J 100:232–242

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Williams DJ (1984) Organic polymeric and non-polymeric materials with large optical non-linearities. Angew Chem Int Ed Engl 23:690–703

    Article  Google Scholar 

  • Wilson SA, Millard A, Lewis A, Loew LM (2014) Monitoring membrane potential with second-harmonic generation. Cold Spring Harb Protoc 2014:643–654

    Article  PubMed  Google Scholar 

  • Yan ECY, Liu Y, Eisenthal KB (1998) New method for determination of surface potential of microscopic particles by second harmonic generation. J Phys Chem B 102:6331–6336

    Article  CAS  Google Scholar 

  • Yan P, Millard AC, Wei M, Loew LM (2006) Unique contrast patterns from resonance-enhanced chiral SHG of cell membranes. J Am Chem Soc 128:11030–11031

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Yan P, Xie A, Wei M-d, Loew LM (2008) Amino(oligo)thiophene-based environmentally sensitive biomembrane chromophores. J Org Chem 73:6587–6594

    Article  CAS  PubMed Central  PubMed  Google Scholar 

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

    Article  CAS  PubMed Central  PubMed  Google Scholar 

Download references

Acknowledgment

We are indebted to the many talented students and colleagues who have participated in the development of SHG imaging in our laboratories over the last 20 years. This work was supported by NIH EB001963.

Author information

Authors and Affiliations

  1. Department of Cell Biology, R. D. Berlin Center for Cell Analysis and Modeling, University of Connecticut Health Center, Farmington, CT, 06030-1507, USA

    Leslie M. Loew

  2. Department of Applied Physics, Hebrew University, Jerusalem, Israel

    Aaron Lewis

Authors
  1. Leslie M. Loew
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Aaron Lewis
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Leslie M. Loew .

Editor information

Editors and Affiliations

  1. Laboratoire Interdisciplinaire de Physique, Grenoble, France

    Marco Canepari

  2. Department of Cellular and Molecular Physiology, Yale University School of Medicine, New Haven, Connecticut, USA

    Dejan Zecevic

  3. L’Institut de rythmologie et mode´lisation cardiaque LIRYC Universite´ de Bordeaux, Bordeaux, France

    Olivier Bernus

Rights and permissions

Reprints and permissions

Copyright information

© 2015 Springer International Publishing Switzerland

About this chapter

Cite this chapter

Loew, L.M., Lewis, A. (2015). Second Harmonic Imaging of Membrane Potential. In: Canepari, M., Zecevic, D., Bernus, O. (eds) Membrane Potential Imaging in the Nervous System and Heart. Advances in Experimental Medicine and Biology, vol 859. Springer, Cham. https://doi.org/10.1007/978-3-319-17641-3_19

Download citation

Publish with us

Policies and ethics

Search

Navigation