In situ fiber-optical monitoring of cytosolic calcium in tissue explant cultures

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.

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