Effects of substrates on the relative immigration activities (RIAs) of epilithic diatoms were examined in Sum River, a tributary of South Han River, Korea. Two hundreds of tile substrates coated with 5% agar were deposited at seven study stations (30 tiles per site) for one month from March 7 to April 7, 2008. Water sampling, physico-chemical factor measurement, and diatom analysis were performed in the field and laboratory. Over the study, major epilithic diatom species were Nitzschia amphibia, Navicula subminuscula, Cymbella minuta in both the natural and artificial substrates. Two dominant species, Nitzschia amphibia and Cymbella minuta often found at seven stations. Among all observed taxa, Nav. subminuscula and Caloneis silicula showed the highest RIA, with the value over 15 throughout the study. Regarding the algal morphology, the biraphe type species showed higher RIA than any other morph, while they comprised over 55% of total biomass, indicating a density effect. Additionally, a significant relationship between RIA and 20-day deposited substrate (p<0.05) showed in two diatom species, Nitzschia fonticola, Gomphonema quadripunctatum, which showed relatively high RIAs. These results indicate that the relative immigration activities of epilithic diatom species can be influenced by algal density in the water and substrate, nutrients, and deposited time at least 20 days.