The present investigation is an attempt to create an optimized protocol for a bactericidal modality of different powers of He-Ne laser radiation to eliminate periodontitis disease causing bacteria from dental plaques. Periodontitis is most prevalent infectious disease of men and caused by a limited number of Gram negative oral bacteria. Porphyromonas gingivalis and Streptococcus sanguis are the important bacteria responsible for periodontitis diseases. Effect on periodontitis disease causing bacteria were produced by the exposure of different powers of He-Ne laser light i.e. 9 mW, 17 mW and 26 mW of red colour of wavelength 632.8 nm in two different periods of time i.e. 10 min. and 20 min. in the presence of dye Methylene blue (MB) used as a photosensitizer. The results have been shown in terms of percentage inhibition of colony forming units (cfu.) of bacteria. This study has shown that maximum inhibition of cfu. were observed in Laser+MB-20 min. exposure time. This inhibition was followed by Laser+MB-10 min., but minimum inhibition was seen in Laser only at 10 min. exposure. In case of effect of methylene alone on the cfu. of bacteria, it was seen that MB have not shown more inhibition of cfu. and it had shown that the no. of cfu. are very similar to that of control. The above observation of the present study was seen in case of every 3 different type of used powers of laser for both the bacteria. Maximum percentage inhibition of cfu. were seen in case of 26mW powers of He-Ne laser, which was 67. 28% to 61.42% for Porphyromonas gingivalis and Streptococcus sanguis respectively. So, increasing the power of laser (safe range for dentistry is 3-30 mW) under conditions shows an increased percentage inhibition of cfu. Thus the present investigation may be a useful adjunct with mechanical debridement in the prevention of recolonization of subgingival lesions by pathogenic microorganisms which are harmful and drug resistant.