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Advanced Time-Correlated Single Photon Counting Applications pp 225–247Cite as

Imaging Cell and Tissue O2 by TCSPC-PLIM

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Part of the book series: Springer Series in Chemical Physics ((CHEMICAL,volume 111))

Abstract

We describe a technique of imaging tissue oxygen using phosphorescence based probes and TCSPC-PLIM method. Included is a brief overview of the significance of biological oxygen imaging, the theory behind the phosphorescence quenching method, the main O2 sensitive probes (mostly intracellular, cell-permeable) and imaging modalities currently available, highlighting their merits and limitations. In the practical part, the live cell microscopy imaging and TCSPC-PLIM hardware and software are described, along with the detailed experimental procedures of preparation of tissue samples, their staining with intracellular O2 probes, acquisition of PLIM images and their processing to produce 2D and 3D maps of O2 concentration. Several examples demonstrate practical use of O2 imaging with different models of mammalian tissue, including cell mono-layers (2D model), multi-cellular spheroids, scaffold structures and tissue slices (3D models). Physiological experiments and multi-parametric analysis of these samples with some other fluorescent imaging probes are also presented.

James Jenkins and Ruslan I. Dmitriev these authors contributed equally to this work.

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Acknowledgments

This work was supported by the Science Foundation Ireland, grant 12/RC/2276, the European Commission FP7 Program, grant FP7-HEALTH-2012-INNOVATION-304842-2.

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Authors and Affiliations

  1. School of Biochemistry and Cell Biology, University College Cork, Cork, Ireland

    James Jenkins

  2. School of Biochemistry and Cell Biology, University College Cork, Western Road, Cork, Ireland

    Ruslan I. Dmitriev

  3. Laboratory of Biophysics and Bioanalysis, School of Biochemistry and Cell Biology, University College Cork, Cork, Ireland

    Dmitri B. Papkovsky

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  1. James Jenkins
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  2. Ruslan I. Dmitriev
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  3. Dmitri B. Papkovsky
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Correspondence to Dmitri B. Papkovsky .

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  1. Becker & Hickl GmbH, Berlin, Germany

    Wolfgang Becker

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Jenkins, J., Dmitriev, R.I., Papkovsky, D.B. (2015). Imaging Cell and Tissue O2 by TCSPC-PLIM. In: Becker, W. (eds) Advanced Time-Correlated Single Photon Counting Applications. Springer Series in Chemical Physics, vol 111. Springer, Cham. https://doi.org/10.1007/978-3-319-14929-5_6

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