OSA's Digital Library

Optics Express

Optics Express

  • Editor: C. Martijn de Sterke
  • Vol. 17, Iss. 2 — Jan. 19, 2009
  • pp: 984–990

Spectral characterization of the voltage-sensitive dye di-4-ANEPPDHQ applied to probing live primary and immortalized neurons

Yu Wang, Gaoshan Jing, Susan Perry, Filbert Bartoli, and Svetlana Tatic-Lucic  »View Author Affiliations

Optics Express, Vol. 17, Issue 2, pp. 984-990 (2009)

View Full Text Article

Enhanced HTML    Acrobat PDF (668 KB)

Browse Journals / Lookup Meetings

Browse by Journal and Year


Lookup Conference Papers

Close Browse Journals / Lookup Meetings

Article Tools



Spectral properties of a recently developed voltage-sensitive dye, di-4-ANEPPDHQ, were characterized as the dye was dissolved in the solvent dimethyl sulfoxide as the stock solution, in Hank's buffered salt solution as the staining solution, and bound to the plasma membrane of primary rat hippocampal neurons and immortalized mouse hypothalamic neurons (GT1-7) in vitro. Their dependence on the local chemical and electrical environment of dye molecules was determined. The excitation and emission peaks are 479nm and 570 nm for the stained primary neurons, and 476nm and 585 nm for the stained immortalized neurons. The excitation and emission bands of the stained GT1-7 neurons, defined as 50% peak intensity, are 429-516nm and 544-648nm, respectively.

© 2009 Optical Society of America

OCIS Codes
(170.0170) Medical optics and biotechnology : Medical optics and biotechnology
(300.6170) Spectroscopy : Spectra

ToC Category:
Medical Optics and Biotechnology

Original Manuscript: November 6, 2008
Revised Manuscript: December 31, 2008
Manuscript Accepted: January 12, 2009
Published: January 13, 2009

Virtual Issues
Vol. 4, Iss. 3 Virtual Journal for Biomedical Optics

Yu Wang, Gaoshan Jing, Susan Perry, Filbert Bartoli, and Svetlana Tatic-Lucic, "Spectral characterization of the voltage-sensitive dye di-4-ANEPPDHQ applied to probing live primary and immortalized neurons," Opt. Express 17, 984-990 (2009)

Sort:  Author  |  Year  |  Journal  |  Reset  


  1. B. Alberts, D. Bray, K. Hopkin, A. Johnson, J. Lewis, M. Raff, K. Roberts, and P. Walter, Essential Cell Biology, 2nd edition, (Garland Science, New York, 2003).
  2. L. B. Cohen and B. M. Salzberg, "Optical measurement of membrane potential," Reviews of Physiology, Biochemistry and Pharmacology (Springer Berlin/Heidelberg), 83, 35-88 (1978). [PubMed]
  3. P. Fromherz, K. H. Dambacher, H. Ephardt, A. Lambacher, C. O. Müller, R. Neigl, H. Schaden, O. Schenk, and T. Vetter, "Fluorescent dyes as probes of voltage transients in neuron membranes," Berichte der Bunsengesellschaft fuer Physikalische Chemie 95, 1333-1345 (1991).
  4. M. Djurisic, M. Zochowski, M. Wachowiak, C. X. Falk, L. B. Cohen, and D. Zecevic, "Optical monitoring of neural activity using voltage-sensitive dyes," Methods Enzymol. 361, 423-451 (2003). [CrossRef] [PubMed]
  5. L. M. Loew, "Potentiometric dyes: imaging electrical activity of cell membranes," Pure Appl. Chem. 68, 1405-1409 (1996). [CrossRef]
  6. L. M. Loew, L. B. Cohen, J. Dix, E. N. Fluhler, V. Montana, G. Salama, and J. Y. Wu, "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. Membrane Bio. 130, 1-10 (1992).
  7. Invitrogen Corporation, "Potential-sensitive ANEP dyes," Data sheet, Revised: 24-March-2006, available at http://probes.invitrogen.com/media/pis/mp01199.pdf, November 5, 2008.
  8. J. A. N. Fisher, J. R. Barchi, C. G. Welle, G.-H. Kim, P. Kosterin, A. L. Obaid, A. G. Yodh, D. Contreras and B. M. Salzberg, "Two-photon excitation of potentiometric probes enables optical recording of action potentials from mammalian nerve terminals in situ," J. Neurophysiol. 99, 1545-1553 (2008). [CrossRef] [PubMed]
  9. A. L. Obaid, L. M. Loew, J. P. Wuskell, B. M. Salzberg, "Novel naphthylstyryl-pyridinium potentiometric dyes offer advantages for neural network analysis," J. Neurosci. Methods 134, 179-190 (2004). [CrossRef] [PubMed]
  10. A. L. Obaid, M. E. Nelson, J. Lindstrom and B. M. Salzberg, "Optical studies of nicotinic acetylcholine receptor subtypes in the guinea-pig," J. Experimental Bio. 208, 2981-3001 (2005). [CrossRef]
  11. Y. Roche, P. Gerbeau-Pissot, B. Buhot, D. Thomas, L. Bonneau, J. Gresti, S Mongrand, J.-M. Perrier-Cornet, and F. Simon-Plas, "Depletion of phytosterols from the plant plasma membrane provides evidence for disruption of lipid rafts," The FASEB Journal 22, 1-12 (2008). [CrossRef]
  12. D. M. Owen, P. M. P. Lanigan, C. Dunsby, I. Munro, D. Grant, M. A. A. Neil, P. M. W. French, and A. I. Magee, "Fluorescence lifetime imaging provides enhanced contrast when imaging the phase-sensitive dye di-4-ANEPPDHQ in model membranes and live cells." Biophys. J. 90, L80 - L82 (2006). [CrossRef] [PubMed]
  13. D. M. Owen, M. A.A. Neil, P. M.W. French and A. I. Magee, "Optical techniques for imaging membrane lipid microdomains in living cells," Semin. Cell Biol. 18, 591-598 (2007). [CrossRef]
  14. L. Jin, A. C. Millard, J. P. Wuskell, X. Dong, D. Wu, H. A. Clark, and L. M. Loew, "Characterization and application of a new optical probe for membrane lipid domains," Biophys. J. 90, 2563-2575 (2006). [CrossRef] [PubMed]
  15. L. Jin, A. C. Millard, J. P. Wuskell, H. A. Clark, and L. M. Loew, "Cholesterol-enriched lipid domains can be visualized by di-4-ANEPPDHQ with linear and nonlinear optics," Biophys. J. 89, L04-L06 (2005). [CrossRef] [PubMed]
  16. B. Leuner, E. Gould, and T. J. Shors, "Is there a link between adult neurogenesis and learning?" Hippocampus 16, 216-224 (2006). [CrossRef] [PubMed]
  17. P. L. Mellon, J. J. Windle, P. C. Goldsmith, C. A. Padula, J. L. Roberts, and R. I. Weiner, "Immortalization of hypothalamic GnRH neurons by genetically targeted tumorigenesis," Neuron 5, 1-10 (1990). [CrossRef] [PubMed]
  18. Z. Liposits, I. Merchenthaler, W. C. Wetsel, J. J. Reid, P. L. Mellon, R. I. Weiner, and A. Negro-Vilar, "Morphological characterization of immortalized hypothalamic neurons synthesizing luteinizing hormone-releasing hormone," Endocrinology 129, 1575-1583 (1991). [CrossRef] [PubMed]
  19. G. J. Brewer, J. R. Torricelli, E. K. Evege, and P. J. Price, "Optimized survival of hippocampal neurons in B27 -supplemented neurobasal, a new serum-free medium combination," J. Neurosci. Res. 35, 567-576 (1993). [CrossRef] [PubMed]

Cited By

Alert me when this paper is cited

OSA is able to provide readers links to articles that cite this paper by participating in CrossRef's Cited-By Linking service. CrossRef includes content from more than 3000 publishers and societies. In addition to listing OSA journal articles that cite this paper, citing articles from other participating publishers will also be listed.


Fig. 1. Fig. 2. Fig. 3.
Fig. 4.

« Previous Article  |  Next Article »

OSA is a member of CrossRef.

CrossCheck Deposited