SPIMTOP · Combined Selective Plane Illumination Microscopy and Optical Projection Tomography for in vivo quantitative imaging of the developing zebrafish vasculature

Recently conceived three-dimensional microscopy techniques are revolutionizing developmental biology and embryology studies, being able to visualize entire organisms in-vivo. Despite increasingly proposed imaging solutions, methods suitable to quantify dynamic and mechanical parameters over a large region of a biological specimen are still lacking. This proposal aims to construct an optical system and a computational framework able to quantitatively study the developing embryonic zebrafish vasculature, looking at the interaction between gene expression and hemodynamic parameters, at high spatial resolution, over a large volume of the organism. The instrument will exploit an advanced version of Optical Projection Tomography, in order to provide a detailed three-dimensional map of the zebrafish cardiovascular system annotated with blood flow and shear stress parameters. Co-registered Selective Plane Illumination Microscopy images will enable the users to observe the expression of fluorescent markers at sub-cellular resolution and to correlate it to the hemodynamic/mechanical parameters. Low photo-bleaching and high acquisition speeds make this multimodal technology ideally suited for extended time-lapse experiments in live embryos. The system will be employed to characterize the development of the embryonic cardiovascular system in normal and mutant zebrafish embryos.The researcher, proponent of this project, will be integrated in a highly multidisciplinary team and will be trained on microscopy and biological aspects of the research, being able to acquire new skills relevant for the development of his optical tomography laboratory and his career.""