[S8.4]: Cell tracking of development using 2‐photon scanning and light sheet imaging

We undertake a Complex System approach to achieve the automated phenomenological reconstruction required for a further theoretical reconstruction of morphogenetic processes multiscale dynamics. Our strategies allow achieving the reconstruction of the cell lineage tree from 3D + time 2-photon laser scanning or light sheet microscopy images of zebrafish early embryogenesis. Our standardized and controlled original algorithmic methods provide measurements with precision and accuracy unattained so far. Spatio-temporal correlations taking into account cell division rate and orientation, cell displacements, cell neighborhood persistence and cell fate along the lineage tree allow a first level of qualitative and predictive modeling of morphogenetic processes underlying gastrulation and brain early embryogenesis. The 4D lineage tree is the framework to further integrate lower level dynamics (molecular and genetic) and higher level features (cell collective behaviors and system biomechanics). Achieving the reconstruction of gene regulation dynamics requires gathering quantitative 4D gene expression data to constrain models parameters and integrate dynamical processes reconstructed at the molecular and cellular level of organization. This is obtained with a combination of QPCR, 3D data from fluorescent in situ hybridization and 3D + time data from live transgenic fish lines. We will discuss the relevance of our strategies in terms of biological systems understanding, prediction and control. Furthermore, complex systems approaches lead biologists to produce data that can only be handled in the context of interdisciplinary interactions. The pioneering infrastructure called “BioEmergences” that we developed in collaboration with the calculation centre of the CNRS and the French portal to the EGEE grid respond to the requirements of next generation e-biology.