Transgenic zebrafish in which the germline is specifically labeled with enhanced green fluorescent protein (eGFP) can be used for continuous observation of germline development during the lifetime, from the primordial germ cells (PGCs) in the early embryo to the gametes in the mature gonad. In this chapter, we describe a procedure for the generation of transgenic fish Tg(piwil1:egfp-UTRnanos3), the sample preparation for live imaging of PGCs, for high-resolution imaging of germ cells in developing gonads, and quantifying PGC numbers. The methods described in this chapter are not only applicable to the study of germ cells, but also provide general advices for researchers who are willing to generate transgenic zebrafish and do observation on live embryos as well as on fixed tissues.
Keywords: Confocal imaging; Germline; Live imaging; Sample preparation; Transgenic fish.