In this paper,optical spectroscopy could uses for medical devices, UV-Vis, photoluminescence(PL) and laser induced fluorescence spectroscopy(LIF) are employed to investigate different concentration of cholesterol(CH). The effect of CH concentration on the emission wavelength of dyes(RdB,Rd6G, Methylene Blue) was investigated by using LIF. It is shown that increasing in CH concentration leads to decrease in the photoluminescence spectrum and increase in the emission wavelength (Red Shif). In the study of the effect of CH concentration on the emission wavelength of Rhodamine6G, increasing in CH concentration in this dye caused red spectral shift and in the two other dyes the spectral shift have not been observed, it could find cholesterol has no direct reaction with these dyes.