Pflügers Archiv

, Volume 445, Issue 3, pp 357–365

Two-photon chloride imaging in neurons of brain slices

  • Nima Marandi
  • Arthur Konnerth
  • Olga Garaschuk
Original Article

DOI: 10.1007/s00424-002-0933-7

Cite this article as:
Marandi, N., Konnerth, A. & Garaschuk, O. Pflugers Arch - Eur J Physiol (2002) 445: 357. doi:10.1007/s00424-002-0933-7

Abstract.

Two-photon laser scanning microscopy has been used successfully for imaging activity-dependent changes of intracellular calcium and sodium levels. Here we introduce a simple technique for two-photon chloride imaging in intact neurons. It involves the use of the membrane-permeable Cl indicator dye MQAE [N-(6-methoxyquinolyl) acetoethyl ester]. Brief incubation with MQAE produced a robust loading of cells in slices from various brain regions including hippocampus, cortex and cerebellum. In contrast to conventional fluorescence measurements using MQAE, two-photon imaging was not affected in a major way by dye bleaching and phototoxic damage. Instead, it allowed prolonged recordings of time-resolved changes in intracellular chloride concentration in somata and dendrites. As an example of an application we imaged GABA-mediated Cl transients in pyramidal cells of cortical and hippocampal slices as well as in cerebellar Purkinje neurons. By combining Cl imaging with the gramicidin-based perforated-patch-clamp technique we showed that changes in MQAE fluorescence are proportional to the magnitudes of GABA-evoked transmembrane Cl fluxes. Thus, MQAE-based two-photon microscopy promises to be a valuable technique for many applications requiring chloride imaging in single cells.

Cl– imaging Dendrite MQAE Two-photon excitation 

Copyright information

© Springer-Verlag 2002

Authors and Affiliations

  • Nima Marandi
    • 1
  • Arthur Konnerth
    • 1
  • Olga Garaschuk
    • 1
  1. 1.Institut für Physiologie, Ludwig-Maximilians Universität München, Pettenkoferstrasse 12, 80336 Munich, Germany

Personalised recommendations