Article abstract
Nature Photonics 3, 412 - 417 (2009)
Published online: 21 June 2009 | doi:10.1038/nphoton.2009.98
Subject Category: Imaging and sensing
Multispectral opto-acoustic tomography of deep-seated fluorescent proteins in vivo
Daniel Razansky1, Martin Distel2, Claudio Vinegoni3, Rui Ma1, Norbert Perrimon4, Reinhard W. Köster2 & Vasilis Ntziachristos1
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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Institute for Biological and Medical Imaging, Technical University of Munich and Helmholtz Center Munich, Ingolstädter Landstra
e 1, D-85764 Neuherberg, Germany
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Institute of Developmental Genetics, Helmholtz Center Munich, Ingolstädter Landstrae 1, D-85764 Neuherberg, Germany
- Center for Systems Biology, Massachusetts General Hospital and Harvard Medical School, Richard B. Simches Research Center, 185 Cambridge Street, Boston, Massachusetts 02114, USA
- Department of Genetics, Harvard Medical School and Howard Hughes Medical Institute, 77 Avenue Louis Pasteur, Boston, Massachusetts 02115, USA
Correspondence to: Daniel Razansky1 e-mail: dr@tum.de
Correspondence to: Vasilis Ntziachristos1 e-mail: v.ntziachristos@tum.de
