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Keywords:

  • cytosolic calcium;
  • cell culture;
  • online monitoring;
  • Calcium Green-5N;
  • fiber-optic;
  • ciliated airway epithelium;
  • ratiometric fluorescence spectroscopy;
  • autofluorescence

Abstract

We present a fluorescence-lifetime based method for monitoring cell and tissue activity in situ, during cell culturing and in the presence of a strong autofluorescence background. The miniature fiber-optic probes are easily incorporated in the tight space of a cell culture chamber or in an endoscope. As a first application we monitored the cytosolic calcium levels in porcine tracheal explant cultures using the Calcium Green-5N (CG5N) indicator. Despite the simplicity of the optical setup we are able to detect changes of calcium concentration as small as 2.5 nM, with a monitoring time resolution of less than 1 s. (© 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim)