The study of the mechanisms underlying both neural development and functioning in vivo is today possible thanks to light sheet fluorescence microscopy (LSFM) techniques, such as Single Plane Illumination Microscopy (SPIM). In this project, we develop a methodology to image developing zebrafish embryos using the SPIM. This allows us to identify neural structures expressing an extracellular matrix protein, F-spondin, and to characterize migration patterns of optic tectum (Tec) neurons. We image zebrafish embryos (transgenic line spon1b:GFP) from 24 to 70 hours post-fertilization (hpf), and we have identified possible primordia of olfactory bulbs (OB) and habenular (Hb) progenitors at 24 hpf. Additionally, we apply image processing and analysis to obtain single cell trajectories and the morphology, to determine the mechanisms that might be important during optic tectum development.