A method for the preparation of Rhodamine 6G immobilized on gold nanoparticles, comprising steps of gold nanoparticle synthesis by citrate reduction method using tetrachloroauric acid and trisodium citrate at 95°C and the colour change to wine red indicated the synthesis completion. The unreacted components and impurities were removed by repeated centrifugation at13000 rpm for 20 min and the pellet containing citrate capped gold nanoparticles were collected. This pellet of citrate capped gold nanoparticles was dissolved in deionized water at a concentration of 18 µM and Rhodamine 6G was added to this solution at 1.7µM for electrostatic conjugation. The solution was stirred using magnetic stirrer for 2 hours to get a stable product of Rhodamine 6G immobilized gold nanoparticles. The product had a size of 20-30 nm and zeta potential of -27 mV, which indicated small size and high stability, respectively. The product has enhanced ROS generating capacity than its individual components and was applied to inhibit the growth of multidrug resistant bacteria and uncontrolled proliferating mammalian cells. The product showed anti-Stokes effect and was also applied for fluorescence and CT imaging.