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Proceedings Paper

Confocal fluorescence lifetime imaging of ion concentrations
Author(s): Hans C. Gerritsen; Renata Sanders; Arie Draaijer
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Paper Abstract

Ion concentrations in biological cells are widely studied with fluorescent probes. The probes have a high selectivity for specific ions and exhibit marked changes in their photophysical properties upon binding ions. The fluorescence decay behavior of the probes in the presence of ions 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 living cells. Here we describe the fluorescence lifetime properties of the free calcium probe CalciumGreen and the pH probe carboxy SNAFL-1. The potential of fluorescence lifetime imaging is illustrated by the imaging of Ca2+ concentrations and pH in single cells. In the case of the emission ratio probe c.SNAFL-1, it was possible to determine the pH in the same cell using both the ratio and the fluorescence lifetime method. It turns out that no cumbersome in vivo calibration procedure is required when c.SNAFL-1 is used for quantitative fluorescence lifetime imaging of pH in single cells.

Paper Details

Date Published: 1 February 1995
PDF: 8 pages
Proc. SPIE 2329, Optical and Imaging Techniques in Biomedicine, (1 February 1995); doi: 10.1117/12.200890
Show Author Affiliations
Hans C. Gerritsen, Univ. of Utrecht (Netherlands)
Renata Sanders, Univ. of Utrecht and TNO Institute of Environmental Sciences (Netherlands)
Arie Draaijer, Univ. of Utrecht and TNO Institute of Environmental Sciences (Netherlands)


Published in SPIE Proceedings Vol. 2329:
Optical and Imaging Techniques in Biomedicine
Hans-Jochen Foth; Aaron Lewis; Halina Podbielska; Michel Robert-Nicoud; Herbert Schneckenburger; Anthony J. Wilson, Editor(s)

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