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Confocal fluorescence lifetime imaging of free calcium in single cells

  • Fluorescence Imaging and Microscopy
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Abstract

Ca2+ concentrations in biological cells are widely studied with fluorescent probes. The probes have a high selectivity for free calcium and exhibit marked changes in their photophysical properties upon binding. The differences in the fluorescent lifetime of the probes can now be used as a contrast mechanism for imaging purposes. This technique can be further exploited for the quantitative determination of ion concentrations within the cells. We describe the use of a fast fluorescence lifetime imaging method in combination with a standard confocal laser scanning microscope for the determination of Ca2+ concentrations in single rat cardiac myocytes using the intensity probe Calcium Green.

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Authors and Affiliations

  1. Department of Molecular Biophysics, Utrecht University, Buys Ballot Laboratory, P.O. Box 80.000, 3508 TA, Utrecht, The Netherlands

    Renata Sanders, Hans C. Gerritsen & Yehudi K. Levine

  2. TNO Institute of Environmental Sciences, P.O. Box 6011, 2600 JA, Delft, The Netherlands

    Renata Sanders, Arie Draaijer & Piet M. Houpt

Authors
  1. Renata Sanders
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  2. Hans C. Gerritsen
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  3. Arie Draaijer
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  4. Piet M. Houpt
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  5. Yehudi K. Levine
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Sanders, R., Gerritsen, H.C., Draaijer, A. et al. Confocal fluorescence lifetime imaging of free calcium in single cells. J Fluoresc 4, 291–294 (1994). https://doi.org/10.1007/BF01881442

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  • DOI: https://doi.org/10.1007/BF01881442

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