With a specific photosensitizer density and light dose,Raman spectroscopy technology is applied to study the photodynamic killing effect to C6 glioma cell which experience different incubation time.In the experiment,5 trail groups are used(one contrast group and four photosensitized group with incubation time of 1,3,6,9 hour).Raman spectroscopy of C6 glioma cell for each group are collected.Results show that both peak intensity and peak position change greatly for nucleic acid and protein spectral lines.there is a little change of peak for spectral lines with 1 hour incubation time.The peak intensity and position change most for spectral lines with 6 hour incubation time. As for the spectral lines with 9 hour incubation;the change reach a flat period.It comes to the conclusion that there is a positive relation between the killing effect and the photosensitizing incubation time, but the relation is not obvious when the incubation time exceeds a certain value.