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Multispectral opto-acoustic tomography of deep-seated fluorescent proteins in vivo

Nature Photonics volume 3, pages 412–417 (2009)Cite this article

Abstract

Fluorescent proteins have become essential reporter molecules for studying life at the cellular and sub-cellular level, re-defining the ways in which we investigate biology. However, because of intense light scattering, most organisms and tissues remain inaccessible to current fluorescence microscopy techniques at depths beyond several hundred micrometres. We describe a multispectral opto-acoustic tomography technique capable of high-resolution visualization of fluorescent proteins deep within highly light-scattering living organisms. The method uses multiwavelength illumination over multiple projections combined with selective-plane opto-acoustic detection for artifact-free data collection. Accurate image reconstruction is enabled by making use of wavelength-dependent light propagation models in tissue. By performing whole-body imaging of two biologically important and optically diffuse model organisms, Drosophila melanogaster pupae and adult zebrafish, we demonstrate the facility to resolve tissue-specific expression of eGFP and mCherrry fluorescent proteins for precise morphological and functional observations in vivo.

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Figure 1: Experimental setup of multispectral opto-acoustic tomography.
Figure 2: Multispectral opto-acoustic imaging of tissue-mimicking phantom containing DsRed-expressing HeLa cells.
Figure 3: Imaging of eGFP distribution in Drosophila melanogaster pupa.
Figure 4: Imaging of mCherry distribution in the vertebral column of an adult zebrafish.
Figure 5: Three-dimensional in vivo imaging through the brain of an adult (six-month-old) mCherry-expressing transgenic zebrafish.

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Acknowledgements

D.R. acknowledges financial support by the Deutsche Forschungsgemeinschaft (DFG) research grant RA 1848/1-1. M.D. is a fellow of the Studienstiftung des deutschen Volkes. R.W.K. is supported by a BioFuture Award Grant (0311889) of the German Ministry for Education and Research (BMBF). We thank R. Jagasia for providing Hela mitodsRed cells.

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Authors and Affiliations

  1. Institute for Biological and Medical Imaging, Technical University of Munich and Helmholtz Center Munich, Ingolstädter Landstraße 1, Neuherberg, D-85764, Germany

    Daniel Razansky, Rui Ma & Vasilis Ntziachristos

  2. Institute of Developmental Genetics, Helmholtz Center Munich, Ingolstädter Landstraße 1, Neuherberg, D-85764, Germany

    Martin Distel & Reinhard W. Köster

  3. Center for Systems Biology, Massachusetts General Hospital and Harvard Medical School, Richard B. Simches Research Center, 185 Cambridge Street, Boston, 02114, Massachusetts, USA

    Claudio Vinegoni

  4. Department of Genetics, Harvard Medical School and Howard Hughes Medical Institute, 77 Avenue Louis Pasteur, Boston, 02115, Massachusetts, USA

    Norbert Perrimon

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  1. Daniel Razansky
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  2. Martin Distel
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  7. Vasilis Ntziachristos
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Correspondence to Daniel Razansky or Vasilis Ntziachristos.

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Razansky, D., Distel, M., Vinegoni, C. et al. Multispectral opto-acoustic tomography of deep-seated fluorescent proteins in vivo. Nature Photon 3, 412–417 (2009). https://doi.org/10.1038/nphoton.2009.98

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