title: 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.
authors.affiliation: Institute of Applied Physics, University of Bern, Sidlerstrasse 5, 3012 Bern, Switzerland.
authors.forename: Manuel
authors.initials: M
authors.lastname: Ryser
authors.forename: Lisa
authors.initials: L
authors.lastname: Künzi
authors.forename: Marianne
authors.initials: M
authors.lastname: Geiser
authors.forename: Martin
authors.initials: M
authors.lastname: Frenz
authors.forename: Jaro
authors.initials: J
authors.lastname: Rička
mesh_terms: D000818:Animals; D002118:Calcium / Q000378:metabolism*; D002138:Calibration; D004396:Coloring Agents / Q000378:metabolism; D003600:Cytosol / Q000378:metabolism*; D004847:Epithelial Cells / Q000166:cytology; D006813:Humidity; D055100:Optical Fibers; D061848:Optical Imaging / Q000295:instrumentation*; D009930:Organic Chemicals / Q000378:metabolism; D013552:Swine; D013696:Temperature; D014132:Trachea / Q000166:cytology
chemical_list: D004396:Coloring Agents; D009930:Organic Chemicals; C071336:calcium green; D002118:Calcium
keywords: Calcium Green-5N; autofluorescence; cell culture; ciliated airway epithelium; cytosolic calcium; fiber-optic; online monitoring; ratiometric fluorescence spectroscopy