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Journal of Fluorescence

, Volume 22, Issue 5, pp 1223–1229 | Cite as

Express Method for Determination of Low Value of Trans-membrane Potential of Living Cells with Fluorescence Probe: Application on Haemocytes at Immune Responses

  • Yuriy I. Glazachev
  • Alexandra D. Semenova
  • Natalia A. Kryukova
  • Irina A. Slepneva
  • Viktor V. Glupov
ORIGINAL PAPER

Abstract

The method for measurement of trans-membrane potential of cell membrane was evaluated for the case of low potential value using fluorescence probe 4-(4-dimethylaminostyryl)-1-methylpyridinium, DSM. The method is based on comparative titration of cells with probe in buffers containing Na+ or K+. The apparent trans-membrane potential obtained with this way is a result of K+-Na+ pump activity. The presented approach allowed measuring the low value of potential with 1–2 mV of accuracy without additional calibration procedures. The method was applied for investigation of potential of cell membrane of haemocytes of Galleria mellonella larvae. The value of potential of intact insect’s haemocytes was found in the range from −10 to −20 mV. The change of potential value of haemocytes was investigated under model immune response and natural envenomation and parasitizing. The obtained deviations of cell membrane potential were in good correlation with changes of activity of main immune reactions, described in literature and obtained by us earlier.

Keywords

Potential sensitive probe K+−Na+ pump Immune response Parasitizing 

Notes

Acknowledgments

This study was supported by Russian Foundation for Basic Research (No. 09-04-01582), SB RAS No 46 and by Ministry of Education and Science of Russian Federation, # 2010–1.1-133-133-030.

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Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  • Yuriy I. Glazachev
    • 1
  • Alexandra D. Semenova
    • 1
  • Natalia A. Kryukova
    • 2
  • Irina A. Slepneva
    • 1
  • Viktor V. Glupov
    • 2
  1. 1.Institute of Chemical Kinetics and CombustionSiberian Branch of the Russian Academy of SciencesNovosibirskRussia
  2. 2.Institute of Systematics and Ecology of AnimalsSiberian Branch of the Russian Academy of SciencesNovosibirskRussia

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