Time-Lapse Microscopy of Brain Development

This chapter introduces some of the techniques that can be used for intravital, noninvasive, time lapse confocal microscopy of zebrafish embryonic brain development. No single experimental approach covers all the questions of brain development. But the diVerent approaches have in common that they all follow the basic line of labeling the embryo and embedding it, followed by data recording and data analysis. Thus, the following sections keep this order, introducing and weighing different known techniques for each experimental step. This chapter illustrates that Zebrafish embryos represent an ideal vertebrate model organism for noninvasive intravital imaging because of their optical clarity, external embryogenesis, and fast development. Many different labeling techniques have been adopted from other model organisms or newly developed to address a wealth of different developmental questions directly inside the living organism. The parallel advancements in the field of optical imaging now observe dynamic processes at the cellular and subcellular resolution. Combined with the repertoire of available surgical and genetic manipulations, zebrafish embryos provide powerful and almost unique possibility to observe the interplay of molecular signals with cellular, morphological, and behavioral changes directly within a living and developing vertebrate organism. Finally, this chapter concludes that a bright future for zebrafish is yet to come, let there be light