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Deep in vivo photoacoustic imaging of mammalian tissues using a tyrosinase-based genetic reporter

Nature Photonics volume 9, pages 239246 (2015) | Download Citation

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Abstract

Photoacoustic imaging allows absorption-based high-resolution spectroscopic in vivo imaging at a depth beyond that of optical microscopy. Until recently, photoacoustic imaging has largely been restricted to visualizing the vasculature through endogenous haemoglobin contrast, with most non-vascularized tissues remaining invisible unless exogenous contrast agents are administered. Genetically encodable photoacoustic contrast is attractive as it allows selective labelling of cells, permitting studies of, for example, specific genetic expression, cell growth or more complex biological behaviours in vivo. In this study we report a novel photoacoustic imaging scanner and a tyrosinase-based reporter system that causes human cell lines to synthesize the absorbing pigment eumelanin, thus providing strong photoacoustic contrast. Detailed three-dimensional images of xenografts formed of tyrosinase-expressing cells implanted in mice are obtained in vivo to depths approaching 10 mm with a spatial resolution below 100 μm. This scheme is a powerful tool for studying cellular and genetic processes in deep mammalian tissues.

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Acknowledgements

This work was funded by the UK Biotechnology Research Council (BBSRC) grant no. BB/I014357/1. Additional funding was provided by the gene-therapy division of the UK NIHR University College London Hospital Biomedical Research Centre. This work was also supported by King's College London and University College London Comprehensive Cancer Imaging Centre, Cancer Research UK and the Engineering and Physical Sciences Research Council (EPSRC), in association with the Medical Research Council and Department of Health, UK, and European Union project FAMOS (FP7 ICT, contract no. 317744). P.B. is funded by an EPSRC Leadership Fellowship and J.L. is funded by an ERC starting grant (281356). The authors thank J. Paterson (UCL Advanced Diagnostics) for assistance with immunohistochemistry, K. Venner for assistance with transmission electron microscopy (TEM) and C. Futter for assistance in interpreting the electron micrographs. H. Dortay (TU Berlin) is thanked for helpful comments on the manuscript and P. Varga (AO Research Institute Davos, Switzerland) for assistance with the use of Amira.

Author information

Author notes

    • Jan Laufer

    Present address: Institut für Optik und Atomare Physik, Technische Universität Berlin, Strasse des 17. Juni 135, Berlin 10623, Germany

    • Amit P. Jathoul
    • , Jan Laufer
    • , Martin A. Pule
    •  & Paul Beard

    These authors contributed equally to this work

Affiliations

  1. Department of Haematology, University College London, Gower Street, London WC1E 6BT, UK

    • Amit P. Jathoul
    • , Arnold R. Pizzey
    • , Brian Philip
    •  & Martin A. Pule
  2. Cancer Institute, University College London, Gower Street, London WC1E 6BT, UK

    • Amit P. Jathoul
    • , Peter Johnson
    • , Arnold R. Pizzey
    • , Brian Philip
    • , R. Barbara Pedley
    •  & Martin A. Pule
  3. Department of Medical Physics & Biomedical Engineering, University College London, Gower Street, London WC1E 6BT, UK

    • Jan Laufer
    • , Olumide Ogunlade
    • , Bradley Treeby
    • , Ben Cox
    • , Edward Zhang
    •  & Paul Beard
  4. Institut für Radiologie, Charité-Universitätsmedizin Berlin, Charitéplatz 1, Berlin 10117, Germany

    • Jan Laufer
  5. UCL Centre for Advanced Biomedical Imaging, University College London, Gower Street, London WC1E 6BT, UK

    • Peter Johnson
    •  & Mark F. Lythgoe
  6. Department of Pathology, University College London, Gower Street, London WC1E 6BT, UK

    • Teresa Marafioti

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Contributions

A.J. carried out molecular cloning, cell preparation, maintenance and analysis, animal work, the design of experiments, in vitro characterizations and in vivo photoacoustic imaging, and assisted with preparation of the manuscript. J.L. undertook the photoacoustic spectroscopy and imaging studies, the reconstruction, processing and analysis of the in vivo images, and assisted with preparation of the manuscript. O.O. contributed to tissue phantom experiments and implemented the cell detection limit study. B.T. and B.C. developed the signal processing, image reconstruction and visualization methods. E.Z. designed and constructed the photoacoustic scanner. P.J. provided cell lines, and helped with in vitro and in vivo imaging and histological analyses. A.P. helped with analysis of cells by flow cytometry and with the general experimental design. RBP carried out the production of virus, and helped with cellular analyses and the use of different iterations of his novel marker gene. T.M. performed immunohistochemistry. M.L. was responsible for invocation of the project, and contributed to experimental planning, motivation, the use of facilities and equipment, experimental focus and editing of the manuscript. R.B. provided cell lines, mice, microscopy and the use of the Home Office Project Licence. M.P. provided gene and vector designs, codon optimization, experimental designs, and directed the overall focus of the work and writing of the manuscript. P.B. directed the photoacoustic imaging component of the project and organized and co-wrote the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Paul Beard.

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DOI

https://doi.org/10.1038/nphoton.2015.22

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