Lifetime imaging of the vital DNA/RNA probe SYTO13 in healthy and apoptotic cells

Marc M.J. van Zandvoort; Wim Engels; Cees J. de Grauw; Hans C. Gerritsen; Dick W. Slaaf

doi:10.1117/12.472113

21 June 2002 Lifetime imaging of the vital DNA/RNA probe SYTO13 in healthy and apoptotic cells

Marc M.J. van Zandvoort, Wim Engels, Cees J. de Grauw, Hans C. Gerritsen, Dick W. Slaaf

Proceedings Volume 4626, Biomedical Nanotechnology Architectures and Applications; (2002) https://doi.org/10.1117/12.472113
Event: International Symposium on Biomedical Optics, 2002, San Jose, CA, United States

Abstract

Of the few vital DNA and RNA probes, the SYTO dyes are the most specific for nucleic acids. However, they show no spectral contrast upon binding to DNA or RNA. We show that two-photon fluorescence lifetime imaging of SYTO13 allows the differential and simultaneous imaging of DNA and RNA in living cells and allows sequential and repetitive assessment. Two-photon main of SYTO13 exhibits a fluorescence lifetime of 3.4 +/- 0.2 ns when associated with nuclear DNA, while bound to RNA its lifetime is 4.1 +/- 0.1 ns. After the induction of apoptosis, clusters of SYTO13 with a fluorescence lifetime of 3.4 +/- 0.2 ns become apparent int eh cytoplasm, identified as mitochondrial DNA on the basis of localization experiments. Upon progression of apoptosis the lifetime of SYTO13 attached to DNA shortens significantly, while no changes are detected in the lifetime of SYTO13 attached to DNA and RNA we started an in vitro study. Here we report the first results of this study. The binding fraction of SYTO13 to DNA increases with increasing amount of DNA (RNA), reaching a plateau for DNA (RNA) concentrations above 0.5 mg/ml. From the spectral measurements we conclude that the natural fluorescence lifetime (tau) ₀ of SYTO13 attached to DNA is 3.7 +/- 0.2 ns, while that of SYTO13 attached to RNA is 4.4 +/- 0.2 ns.

Citation Download Citation

Marc M.J. van Zandvoort, Wim Engels, Cees J. de Grauw, Hans C. Gerritsen, and Dick W. Slaaf "Lifetime imaging of the vital DNA/RNA probe SYTO13 in healthy and apoptotic cells", Proc. SPIE 4626, Biomedical Nanotechnology Architectures and Applications, (21 June 2002); https://doi.org/10.1117/12.472113

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