Gastrulation is a complex set of cellular rearrangements that establish the overall shape of the body plan during development. In addition to being an essential and fascinating aspect of development, the cells of the gastrulating zebrafish embryo also provide an ideal in vivo system to study the interplay of cell polarity and movement in a native 3D environment. During gastrulation, zebrafish mesodermal cells undergo a series of conversions from initial non-polarized amoeboid cell movements to more mesenchymal and finally highly polarized and intercalative cell behaviors. Many of the cellular behavior changes of these cells are under the control of the RhoA pathway, which in turn is regulated by many signals, including non-canonical Wnts. The goal of this chapter is to provide researchers with the necessary protocols to examine changes in cell polarity and movement in the developing zebrafish embryo.