Stable isotope of individual fluid inclusion has great significance in researching the formation of rock and mineral paleo-fluid, geology of ore deposits, oil/gas mineral deposits and geodynamics of tectonic evolution. Micro-Laser Raman analysis is a method for stable isotopes in single fluid inclusions. This study proposed a method to calculate δ13C value with Micro-Laser Raman spectroscopy. 12CO2 and 13CO2 mixed with N2 at various molar fraction ratios respectively through self-devised experimental apparatus. The determination of Raman parameters provides theoretical basis for calculating the δ13C value. Based on the study of principles and feasibility of the method of laser Raman spectroscopy, a series of known molar fractions of 12CO2/13CO2 binary mixtures as artificial CO2 fluid inclusions have been prepared. The artificial CO2 fluid inclusions and CO2 gas from Shengli Oil-field have been analyzed with Micro-Laser Raman analysis method. And the molar fraction ratio N13/N12 is related to Raman parameters, the value of δ13C could be calculated by formula δ13C=［(C13/C12)sample/(C13/C12)PDB-1］×1 000‰. The δ13C value of CO2 gas from Shengli Oil-field is -5.6‰ and -5.318 ‰ by using mass spectrometry and Micro-Laser Raman analysis method, respectively. The results demonstrate that it builds up a way to determine δ13C Value of CO2 with Micro-Laser Raman Spectroscopy.