Multispectral opto acoustic tomography of deep seated fluorescent proteins in vivo英文文献.pdfVIP

ARTICLES PUBLISHED ONLINE: 21 JUNE 2009 | DOI: 10.1038/NPHOTON.2009.98 Multispectral opto-acoustic tomography of deep-seated fluorescent proteins in vivo 1 2 3 1 4 Daniel Razansky *, Martin Distel , Claudio Vinegoni , Rui Ma , Norbert Perrimon , ¨ 2 and Vasilis Ntziachristos1 Reinhard W. Koster * 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. luorescence protein (FP) imaging is increasingly being used as a volumes over long periods of time15. Non-invasive high-resolution key technology allowing the comprehension of complex and