The purpose of this study was to investigate the optical properties, absorption coefficient (μ a ) scattering coefficient (μ s ) and refractive indices, (n) of HeLa cell line in a suspension of 2% minimum essential medium (MEM) at two different (632.8 and 532.0 nm) wave lengths of laser light. Optical properties were determined with Kubelka Munk Model (KMM) and refractive index measurement was made through minimum angle of deviation method (MAD). We reported μ a = 8.643 ± 0.187 and 2.348 ± 0.249 cm -1 and μ s = 5.609 ± 0.287 and 88.166 ± 2.833 cm -1 at 632.8 and 532.0 nm, respectively. Refractive index was found to be 1.332 and 1.312 at 632.8 nm and 532.0 nm, respectively. The discussed results provide a route of information for clinical diagnosis, therapeutic application and dosimetry studies in HeLa and other cell lines.