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In vivo optical molecular imaging and analysis in mice using dorsal window chamber models applied to hypoxia, vasculature and fluorescent reporters

Nature Protocols volume 6, pages 1355–1366 (2011)Cite this article

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Abstract

Optical techniques for functional imaging in mice have a number of key advantages over other common imaging modalities such as magnetic resonance imaging, positron emission tomography or computed tomography, including high resolution, low cost and an extensive library of available contrast agents and reporter genes. A major challenge to such work is the limited penetration depth imposed by tissue turbidity. We describe a window chamber technique by which these limitations can be avoided. This facilitates the study of a wide range of processes, with potential endpoints including longitudinal gene expression, vascular remodeling and angiogenesis, and tumor growth and invasion. We further describe several quantitative imaging and analysis techniques for characterizing in vivo fluorescence properties and functional endpoints, including vascular morphology and oxygenation. The procedure takes ∼2 h to complete, plus up to several weeks for tumor growth and treatment procedures.

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Figure 1: Window chamber.
Figure 2: Equipment and surgical tools.
Figure 3: Imaging mount.
Figure 4: Surgical platform preparation.
Figure 5: Preparing the mouse for surgery.
Figure 6: Securing the dorsal skin fold.
Figure 7: Tumor cell inoculation.
Figure 8: Functional imaging.
Figure 9: Spectral decomposition.

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Acknowledgements

We thank the US Department of Defense Breast Cancer Research Program (Postdoctoral Fellowship W81XWH-07-1-0355 to G.M.P.; Predoctoral Traineeship Award W81XWH-08-BCRP-PREDOC to A.N.F.), the US National Institutes of Health (R01CA40355 to M.W.D.) and the University of Virginia Cancer Center (James and Rebecca Craig Foundation and the NCI Cancer Center Support Grant P30 CA44579 to C.L.F.) for support for this research. We also thank the University of Virginia NanoSTAR Institute for supporting early efforts to develop and test boron nanoparticles as biomedical imaging agents.

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Authors and Affiliations

  1. Department of Radiation Oncology, Duke University Medical Center, Durham, North Carolina, USA

    Gregory M Palmer, Siqing Shan, Gabi Hanna & Mark W Dewhirst

  2. Department of Biomedical Engineering, Duke University, Durham, North Carolina, USA

    Andrew N Fontanella & Mark W Dewhirst

  3. Department of Chemistry, University of Virginia, Charlottesville, Virginia, USA

    Guoqing Zhang & Cassandra L Fraser

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  1. Gregory M Palmer
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  2. Andrew N Fontanella
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  6. Cassandra L Fraser
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  7. Mark W Dewhirst
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Contributions

G.M.P and A.N.F. wrote the manuscript, collected the data presented and prepared the figures. S.S. and M.W.D. assisted in the design of the content of the manuscript and revisions. G.H. assisted in the production of figures and revision of the manuscript. C.L.F. and G.Z. assisted in the development of boron nanoparticle techniques and manuscript revision.

Corresponding author

Correspondence to Mark W Dewhirst.

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Competing interests

C.L.F. has financial interest in Luminesco, Inc., which is commercializing the BNP technology.

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Palmer, G., Fontanella, A., Shan, S. et al. In vivo optical molecular imaging and analysis in mice using dorsal window chamber models applied to hypoxia, vasculature and fluorescent reporters. Nat Protoc 6, 1355–1366 (2011). https://doi.org/10.1038/nprot.2011.349

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