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Advanced Time-Correlated Single Photon Counting Applications pp 189–211Cite as

Determination of Intracellular Chloride Concentrations by Fluorescence Lifetime Imaging

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Part of the book series: Springer Series in Chemical Physics ((CHEMICAL,volume 111))

Abstract

Fluorescence microscopy with membrane-permeable ion-sensitive fluorophores allows the non-invasive determination of intracellular ion concentrations. Chloride is the major anion of intra- and extracellular fluids influencing a great number of physiological processes. The dysfunction of chloride transporters and channels leads to disturbance in chloride homeostasis that can result in diseases of different parts of the body. The different existing chloride sensitive fluorophores and their usefulness in fluorescence lifetime imaging are put forward in this chapter. Fluorescence lifetime imaging of a chloride sensitive quinolinium dye (MQAE) has been established as an elegant tool to determine intracellular chloride concentrations of different cells in living biological tissue. Details of the experimental procedure are described and two case studies—chloride transport by a chloride transporter (KCC2) across the cell membrane and determination of intracellular chloride concentration in glia cells (EAAT1-positive astrocytes)—are presented.

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  1. Institute of Complex Systems 4 (ICS-4, Cellular Biophysics), Forschungszentrum Jülich, 52428, Jülich, Germany

    Thomas Gensch, Verena Untiet, Arne Franzen, Peter Kovermann & Christoph Fahlke

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  1. Thomas Gensch
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  2. Verena Untiet
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  3. Arne Franzen
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  4. Peter Kovermann
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  5. Christoph Fahlke
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Correspondence to Thomas Gensch .

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  1. Becker & Hickl GmbH, Berlin, Germany

    Wolfgang Becker

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Gensch, T., Untiet, V., Franzen, A., Kovermann, P., Fahlke, C. (2015). Determination of Intracellular Chloride Concentrations by Fluorescence Lifetime Imaging. In: Becker, W. (eds) Advanced Time-Correlated Single Photon Counting Applications. Springer Series in Chemical Physics, vol 111. Springer, Cham. https://doi.org/10.1007/978-3-319-14929-5_4

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