Fluorescence lifetime imaging of oxygen in living cells

The usefulness of the fluorescent probe ruthenium tris(2,2′-dipyridyl) dichloride hydrate (RTDP) for the quantitative imaging of oxygen in single cells was investigated utilizing fluorescence lifetime imaging. The results indicate that the fluorescence behavior of RTDP in the presence of oxygen can...

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Bibliographic Details
Published inJournal of fluorescence Vol. 7; no. 1; pp. 11 - 15
Main Authors Gerritsen, H. C., Sanders, R., Draaijer, A., Ince, C., Levine, Y. K.
Format Journal Article
LanguageEnglish
Published New York Springer Nature B.V 1997
Subjects
Dichlorides
Fluorescent indicators
Imaging
Ion concentration
Macrophages
Oxygen
Quenching
Ruthenium
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Summary:The usefulness of the fluorescent probe ruthenium tris(2,2′-dipyridyl) dichloride hydrate (RTDP) for the quantitative imaging of oxygen in single cells was investigated utilizing fluorescence lifetime imaging. The results indicate that the fluorescence behavior of RTDP in the presence of oxygen can be described by the Stem-Volmer equation. This shows that fluorescence quenching by oxygen is a dynamic quenching process. In addition, it was demonstrated that the fluorescence lifetime of RTDP is insensitive to pH, ion concentration, and cellular contents. This implies that a simple calibration procedure in buffers can be used to quantify oxygen concentrations within cells. First fluorescence imaging experiments on J774 macrophages show a nonuniform fluorescence intensity and a uniform fluorescence lifetime image. This indicates that the RTDP is heterogeneously partitioned throughout the cells, while the oxygen concentration is constant.
ISSN:1053-0509
1573-4994
DOI:10.1007/BF02764572