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Probing intracellular oxygen by quenched phosphorescence lifetimes of nanoparticles containing polyacrylamide-embedded [Ru(dpp(SO3Na)2)3]Cl2

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Abstract

Methods for measuring O2 within living cells that rely on luminescent probes are hampered by several factors: local conditions of hydrophobicity, pH, ionic composition, dielectric constant, and photobleaching by free radical species. Use of a polymer-embedded luminophore should minimize these problems. Here we use a Ru(ii) coordination complex embedded within 45 nm hydrodynamic diameter nanoparticles, and demonstrate that both phosphorescence intensity and lifetimes are O2-sensitive, both in aqueous suspensions and intracellularly (e.g. 4.06 versus 1.55 microseconds under anaerobic or aerobic conditions, respectively). Electroporation is necessary for incorporation of the nanoparticles into yeasts: it is more effective with the fission yeast, Schizosaccharomyces pombe, than for the budding yeast, Saccharomyces cerevisiae. However, electroporation was not required for particle uptake into a cultured human cell-line (mammary adenosarcoma MCF-7), although the intracellular distribution of the probe is more general to intracellular compartments when electroporation is employed. These procedures did not compromise vitality of cells over periods of 6 h, as judged by retention of structural characteristics evident in Nomarski interference or confocal microscopy images. Spatial resolution of intracellular structures defined by nanoparticle phosphorescence intensity imaging indicates potential usefulness of the application of lifetime imaging techniques for mapping of intracellular O2 distributions.

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Authors and Affiliations

  1. School of Chemistry, Cardiff University, Cardiff, Wales, CF10 3AT, UK

    Michael P. Coogan & Simon J. A. Pope

  2. Cardiff School of Biosciences, Main Building, Museum Avenue, Cardiff, Wales, CF10 3AT, UK

    Jonathan B. Court, Victoria L. Gray, Siôn H. Lloyd, Coralie O. Millet & David Lloyd

  3. Confocal Microscopy Unit, Cardiff School of Biosciences, Life Sciences Building, Cardiff, Wales, CF10 3US, UK

    Anthony J. Hayes

Authors
  1. Michael P. Coogan
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  2. Jonathan B. Court
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  3. Victoria L. Gray
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  4. Anthony J. Hayes
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  5. Siôn H. Lloyd
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  6. Coralie O. Millet
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  7. Simon J. A. Pope
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  8. David Lloyd
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Corresponding author

Correspondence to David Lloyd.

Additional information

To the memory of Gregorio Weber (1916–1997), who made seminal advances in fluorescence techniques and taught one of us (DL) at the Biochemistry Department, University of Sheffield, 1958-1961. See http://www.cf.ac.uk/biosi/staffinfo/lloyd/weber/index.html

Electronic supplementary information (ESI) available: Data fits for lifetime decay measurements. See DOI: 10.1039/b9pp00071b

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Coogan, M.P., Court, J.B., Gray, V.L. et al. Probing intracellular oxygen by quenched phosphorescence lifetimes of nanoparticles containing polyacrylamide-embedded [Ru(dpp(SO3Na)2)3]Cl2. Photochem Photobiol Sci 9, 103–109 (2010). https://doi.org/10.1039/b9pp00071b

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  • DOI: https://doi.org/10.1039/b9pp00071b

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