The indentation behavior of Ni-based single crystal superalloy (NBSX) considering the crystal orientation effect is studied by experiment and crystal plasticity simulation. Pyramid and cone indenters are used to conduct the indentation tests of NBSX. Hardness values under different crystal orientations are obtained. The pile-up modes are observed by the scanning electron microscope (SEM). Electron Backscattered Diffraction (EBSD) analysis is carried out and the orientation deviation is qualified by the kernel averaged misorientation (KAM) and geometrically necessary dislocation (GND) density distribution maps. Crystal plasticity finite element (CPFE) simulations are conducted the pile-up modes, dominant slip system patterns and GND density distributions. The experiment and simulation reveal the significant effect of crystal orientation on the indentation behavior. The indentation experiment, EBSD analysis and CPFE simulation achieve good agreeement.