Confocal micro-Raman spectroscopy is employed to study the diffusion process of dimethyl sulfoxide (DMSO) in porcine skin optical clearing. The variation of DMSO concentration with time at different depths of the skin was obtained and then the DMSO diffusion coefficient with the passive diffusion model was calculated. Results show that it has a significant difference at different depths of the skin. Also, the DMSO concentration with the depth at different times was obtained and the same method was used to find the change law of the DMSO diffusion coefficient. Results indicate that it also changes with the treatment time. The experimental results are consistent with the theoretical model in a previous study. The current results demonstrate that Raman spectroscopy has the ability to quantitatively monitor the process of optical clearing.