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Maps of in vivo oxygen pressure with submillimetre resolution and nanomolar sensitivity enabled by Cherenkov-excited luminescence scanned imaging

Nature Biomedical Engineering volume 2, pages 254–264 (2018)Cite this article

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Abstract

Low signal-to-noise ratios and limited imaging depths restrict the ability of optical-imaging modalities to detect and accurately quantify molecular emissions from tissue. Here, by using a scanning external X-ray beam from a clinical linear accelerator to induce Cherenkov excitation of luminescence in tissue, we demonstrate in vivo mapping of the oxygenation of tumours at depths of several millimetres, with submillimetre resolution and nanomolar sensitivity. This was achieved by scanning thin sheets of the X-ray beam orthogonally to the emission-detection plane, and by detecting the signal via a time-gated CCD camera synchronized to the radiation pulse. We also show with experiments using phantoms and with simulations that the performance of Cherenkov-excited luminescence scanned imaging (CELSI) is limited by beam size, scan geometry, probe concentration, radiation dose and tissue depth. CELSI might provide the highest sensitivity and resolution in the optical imaging of molecular tracers in vivo.

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Fig. 1: Excitation fluence decrease with depth into tissue for normal fluorescence imaging versus CELSI; the temporal sequence and molecular probe characteristics.
Fig. 2: Radiation beam shape configuration and region of the tissue where Cherenkov light is generated affects contrast to noise measured.
Fig. 3: The geometry of the imaging camera relative to the X-ray beam entrance position affects image contrast.
Fig. 4: Contrast-to-background ratio is affected by the concentration and depth of the object, and the radiation dose used in scanning.
Fig. 5: Source and detector placement affects the reconstructed CELSI images.
Fig. 6: The spatial resolution of CELSI is below 1 mm down to depths in tissue of 25 mm.
Fig. 7: Animal phantom tomography and in vivo validation of the luminescence yield.
Fig. 8: In vivo imaging of \({\boldsymbol{p}}_{{\mathbf{O}}_{\mathbf{2}}}\) in subcutaneous breast adenocarcinomas.

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Acknowledgements

This work has been funded by the Congressionally Directed Medical Research Program for Breast Cancer Research Program, US Army USAMRAA contract W81XWH-16-1-0004 and National Institutes of Health research grants R01 EB024498 and R01 EB018464.

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Authors and Affiliations

  1. Thayer School of Engineering, Dartmouth College, Hanover, NH, USA

    Brian W. Pogue, Ethan P. LaRochelle, Petr Bruža, Rongxiao Zhang, Jennifer R. Shell, Scott C. Davis & David J. Gladstone

  2. Norris Cotton Cancer Center, Dartmouth-Hitchcock Medical Center, Lebanon, NH, USA

    Brian W. Pogue, Scott C. Davis, David J. Gladstone & Lesley A. Jarvis

  3. Faculty of Information Technology, Beijing University of Technology, Beijing, China

    Jinchao Feng

  4. Key Laboratory of OptoElectronic Science and Technology for Medicine of Ministry of Education, Fujian Provincial Key Laboratory of Photonics Technology, Fujian Normal University, Fuzhou, China

    Huiyun Lin

  5. School of Computer Science, University of Birmingham, Birmingham, UK

    Hamid Dehghani

  6. Department of Biochemistry and Biophysics, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA

    Sergei A. Vinogradov

  7. Department of Medicine, Geisel School of Medicine, Dartmouth College, Hanover, NH, USA

    David J. Gladstone & Lesley A. Jarvis

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  1. Brian W. Pogue
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Contributions

B.W.P. conceived the study, supervised all aspects of the work and drafted the manuscript; J.F., H.L., P.B., E.P.L., R.Z. and J.R.S. each completed measurements and data analysis as well as designed the experiments, wrote initial parts of the manuscript, and edited the entire manuscript. H.D. and S.C.D. helped design and analyse the tomography work with J.F., and each edited the manuscript. S.A.V. provided the molecular probe, provided advice on experimental design and data analysis and edited the manuscript. D.J.G. and L.A.J. each contributed advice on radiotherapy design and data interpretation, as well as edited the manuscript.

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Correspondence to Brian W. Pogue.

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Supplementary information

Supplementary Information

Supplementary discussion, methods, figures, tables and video captions.

Reporting Summary

Supplementary Video 1

Three-dimensional view of a xenograft tumour imaged by CELSI.

Supplementary Video 2

Real-time scanned acquisition of CELSI data in vivo.

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Pogue, B.W., Feng, J., LaRochelle, E.P. et al. Maps of in vivo oxygen pressure with submillimetre resolution and nanomolar sensitivity enabled by Cherenkov-excited luminescence scanned imaging. Nat Biomed Eng 2, 254–264 (2018). https://doi.org/10.1038/s41551-018-0220-3

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