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



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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Figure 1: Photoacoustic (PA) scanner for imaging mice in vivo.
Figure 2: Construction and in vitro optical and PA characteristics of Tyr-expressing cells.
Figure 3: In vivo PA images of Tyr-expressing K562 cells after subcutaneous injection into the flank of a nude mouse (λex = 600 nm).
Figure 4: In vivo PA images of Tyr-expressing 293T cells acquired at different times post-innoculation, illustrating cell population growth (λex = 640 nm).
Figure 5: Deep tissue imaging of Tyr-expressing K562 cells (λex = 680 nm).


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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




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.

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Correspondence to Paul Beard.

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The authors declare no competing financial interests.

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Jathoul, A., Laufer, J., Ogunlade, O. et al. Deep in vivo photoacoustic imaging of mammalian tissues using a tyrosinase-based genetic reporter. Nature Photon 9, 239–246 (2015).

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