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Time-domain fluorescence lifetime imaging for intracellular pH sensing in living tissues

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Abstract

pH sensing in living cells represents one of the most prominent topics in biochemistry and physiology. In this study we performed one-photon and two-photon time-domain fluorescence lifetime imaging with a laser-scanning microscope using the time-correlated single-photon counting technique for imaging intracellular pH levels. The suitability of different commercial fluorescence dyes for lifetime-based pH sensing is discussed on the basis of in vitro as well of in situ measurements. Although the tested dyes are suitable for intensity-based ratiometric measurements, for lifetime-based techniques in the time-domain so far only BCECF seems to meet the requirements of reliable intracellular pH recordings in living cells.

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Acknowledgment

This work was funded by the German Federal Department for Science and Education BMBF (IP 03/517) and the German Research Foundation DFG. We thank Prof. B. Walz (Dept. of Animal Physiology, University of Potsdam) for the generous supply of cockroaches.

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Authors and Affiliations

  1. Institute of Chemistry, University of Potsdam, Karl-Liebknecht-Str. 24-25, 14476, Potsdam-Golm, Germany

    Carsten Hille, Maik Berg, Lena Bressel, Dorit Munzke, Philipp Primus, Hans-Gerd Löhmannsröben & Carsten Dosche

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  1. Carsten Hille
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  2. Maik Berg
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  3. Lena Bressel
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  4. Dorit Munzke
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  6. Hans-Gerd Löhmannsröben
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  7. Carsten Dosche
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Correspondence to Carsten Dosche.

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Hille, C., Berg, M., Bressel, L. et al. Time-domain fluorescence lifetime imaging for intracellular pH sensing in living tissues. Anal Bioanal Chem 391, 1871–1879 (2008). https://doi.org/10.1007/s00216-008-2147-0

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  • DOI: https://doi.org/10.1007/s00216-008-2147-0

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