The grain size of prior austenite has a distinct influence on the microstructure and final mechanical properties of steels. Thus, it is significant to clearly reveal the grain boundaries and therefore to precisely characterize the grain size of prior austenite. For NiCrMoV rotor steels quenched and tempered at high temperature, it is really difficult to display the grain boundaries of prior austenite clearly, which limits a further study on the correlation between the properties and the corresponding microstructure. In this paper, an effective etchant was put forward and further optimized. Experimental results indicated that this agent was effective to show the details of grain boundaries, which help analyze fatigue crack details along the propagation path. The optimized corrosion agent is successful to observe the microstructure characteristics and expected to help analyze the effect of microstructure for a further study on the mechanical properties of NiCrMoV rotor steels used in the field of nuclear power.