Analytical and Bioanalytical Chemistry

, Volume 404, Issue 3, pp 785–792 | Cite as

Potential-modulated fluorescence spectroscopy of zwitterionic and dicationic membrane-potential-sensitive dyes at the 1,2-dichloroethane/water interface

  • Toshiyuki OsakaiEmail author
  • Tatsuya Yoshimura
  • Daichi Kaneko
  • Hirohisa Nagatani
  • Sang-Hyun Son
  • Yutaka Yamagishi
  • Koji Yamada
Original Paper


The previously introduced technique of potential-modulated fluorescence (PMF) spectroscopy was used to study the potential-induced fluorescence change of some different dyes at the polarized 1,2-dichloroethane (DCE)/water (W) interface. A zwitterionic dye (POLARIC 488PPS) showed a PMF response similar to that for the previously studied dye (di-4-ANEPPS) with the same ionic state, and the PMF response was likewise explained by the potential-dependent reorientation of the dye at the DCE/W interface. Though a monocationic dye (POLARIC 488PM) showed no distinct PMF signal, a dicationic dye (di-2-ANEPEQ) showed two relatively weak but detectable PMF signals at lower and higher potential. It has thus been found that the ionic state of a potential-sensitive dye strongly influences the potential-induced reorientation of the dye at the interface and consequently its PMF response. These results support the reorientation/solvatochromic mechanism proposed for “slow” dyes but do not necessarily exclude the electrochromic mechanism proposed for “fast” dyes. PMF spectroscopy would provide useful information on the design of slow dyes for the measurement of the resting potential of cell membranes


Potential-dependent reorientation of a zwitterionic membrane-potential-sensitive dye (POLARIC 488PPS) at the oil/water interface and its potential-modulated fluorescence signal (inset)


Membrane-potential-sensitive dye POLARIC™ Liquid/liquid interface Potential modulated fluorescence 



T. Osakai, T. Yoshimura, and D. Kaneko acknowledge Dr. Shunji Kasahara and Dr. Seiji Akimoto of Kobe University for their helpful pieces of advice in the spectroscopic measurements. We also thank Goryo Chemical, Inc. for supplying POLARIC™ probes.

Supplementary material

216_2012_6199_MOESM1_ESM.pdf (896 kb)
ESM 1 (PDF 895 kb)


  1. 1.
    Waggoner A (1979) Optical probes of membrane potential. J Membr Biol 27:317Google Scholar
  2. 2.
    Plášek J, Sigler K (1996) Slow fluorescent indicators of membrane potential: a survey of different approaches to probe response analysis. J Photochem Photobiol B 33:101Google Scholar
  3. 3.
    Loew LM (1996) Potentiometric dyes: imaging electrical activity of cell membranes. Pure Appl Chem 68:1405Google Scholar
  4. 4.
    Fluhler E, Burnham VG, Loew LM (1985) Spectra, membrane binding, and potentiometric responses of new charge shift probes. Biochemistry 24:5749Google Scholar
  5. 5.
    Montana V, Farkas DL, Loew LM (1989) Dual-wavelength ratiometric fluorescence measurements of membrane potential. Biochemistry 28:4536Google Scholar
  6. 6.
    Clark RJ, Zouni A, Holzwarth JF (1995) Voltage sensitivity of the fluorescent probe RH421 in a model membrane system. Biophys J 68:1406Google Scholar
  7. 7.
    Müller W, Windisch H, Tritthart HA (1986) Fluorescent styryl dyes applied as fast optical probes of cardiac action potential. Eur Biophys J 14:103Google Scholar
  8. 8.
    Osakai T, Sawada J, Nagatani H (2009) Potential-modulated fluorescence spectroscopy of the membrane potential-sensitive dye di-4-ANEPPS at the 1,2-dichloroethane/water interface. Anal Bioanal Chem 395:1055Google Scholar
  9. 9.
    Nagatani H, Iglesias RA, Fermín DJ, Brevet PF, Girault HH (2000) Adsorption behavior of charged zinc porphyrins at the water/1,2-dichloroethane interface studied by potential modulated fluorescence spectroscopy. J Phys Chem B 104:6869Google Scholar
  10. 10.
    Nagatani H, Fermín DJ, Girault HH (2001) A kinetic model for adsorption and transfer of ionic species at polarized liquid|liquid interfaces as studied by potential modulated fluorescence spectroscopy. J Phys Chem B 105:9463Google Scholar
  11. 11.
    Nagatani H, Suzuki S, Fermín DJ, Girault HH, Nakatani K (2006) Interfacial behavior of sulforhodamine 101 at the polarized water/1,2-dichloroethane interface studied by spectroelectrochemical techniques. Anal Bioanal Chem 386:633Google Scholar
  12. 12.
    Nagatani H, Ozeki T, Osakai T (2006) Direct spectroelectrochemical observation of interfacial species at the polarized water/1,2-dichloroethane interface by ac potential modulation technique. J Electroanal Chem 588:99Google Scholar
  13. 13.
    Osakai T, Yamada H, Nagatani H, Sagara T (2007) Potential-dependent adsorption of amphoteric rhodamine dyes at the oil/water interface as studied by potential-modulated fluorescence spectroscopy. J Phys Chem C 111:9480Google Scholar
  14. 14.
    Son SH, Abe Y, Yuasa M, Yamagishi Y, Sakai N, Ayabe T, Yamada K (2011) A systematic analysis of aromatic heterocyclic rings in solvatochromic fluorophores. Chem Lett 40:378Google Scholar
  15. 15.
    Son SH, Yamagishi Y, Tani M, Yuasa M, Yamada K (2011) Spectral shifts of the environment-sensitive fluorophore POLARIC in heterogeneous interfaces. Chem Lett 40:989Google Scholar
  16. 16.
    Bard AJ, Inzelt G, Scholz F (eds) (2008) Electrochemical dictionary. Springer, Berlin, pp 284–285Google Scholar
  17. 17.
    Osakai T, Kakutani T, Senda M (1984) A.c. polarographic study of ion transfer at the water/nitrobenzene interface. Bull Chem Soc Jpn 57:370Google Scholar
  18. 18.
    Kakutani T, Senda M (1979) Theory of a.c. polarization and a.c. polarography and voltammetry of surface redox reaction. Bull Chem Soc Jpn 52:3236Google Scholar

Copyright information

© Springer-Verlag 2012

Authors and Affiliations

  • Toshiyuki Osakai
    • 1
    Email author
  • Tatsuya Yoshimura
    • 1
  • Daichi Kaneko
    • 1
  • Hirohisa Nagatani
    • 2
  • Sang-Hyun Son
    • 3
  • Yutaka Yamagishi
    • 3
  • Koji Yamada
    • 3
  1. 1.Department of Chemistry, Graduate School of ScienceKobe UniversityKobeJapan
  2. 2.Faculty of Chemistry, Institute of Science and EngineeringKanazawa UniversityKanazawaJapan
  3. 3.Section of Materials Science, Faculty of Environmental Earth ScienceHokkaido UniversitySapporoJapan

Personalised recommendations