The reflectance spectrum has been widely adopted to extract diagnosis information of human tissue because it possesses the advantages of noninvasive and rapidity. The external pressure brought by fiber optic probe may influence the accuracy of measurement. In this paper, a systematic study is focused on the effects of probe pressure on intrinsic changes of water and scattering particles in tissue. According to the biphasic nonlinear mixture model, the pressure modulated reflectance spectrum of both in vitro and in vivo tissue is measured and processed with second-derivation. The results indicate that the variations of bulk and bonded water in tissue have a nonlinear relationship with the pressure. Differences in tissue structure and morphology contribute to site-specific probe pressure effects. Then the finite element (FEM) and Monte Carlo (MC) method is employed to simulate the deformation and reflectance spectrum variations of tissue before and after compression. The simulation results show that as the pressure of fiber optic probe applied to the detected skin increased to 80 kPa, the effective photon proportion form dermis decreases significantly from 86% to 76%. Future designs might benefit from the research of change of water volume inside the tissue to mitigate the pressure applied to skin.