In order to assess the survival success of microphytoplankton species in ship ballast water, we examined microphytoplankton diversity from international commercial ships berthed at Ulsan and Pyeongtaek Ports, Korea, and also subjected them to laboratory studies. The ages of ballast water in each ship ranged from 1 to 365 days. Vessels originated from coastal China (Weihai, Lianyunsang and Shanghai), Chile, and from the Yellow and Pacific Oceans. The numbers of species and phytoplankton standing crops in uploaded ballast water were significantly related to the age of ballast water. The most diverse taxonomic group was diatoms. In the laboratory study, the value of in vivo fluorescence in M/V Spring Lyra gradually increased with increasing nutrients such as nitrate and phosphate. Phytoplankton in new (9 days), medium (31 days) and old (365 days) ballast water successfully survived under typical nutrient condition of port water and F/2 medium at $15^{circ}C$ and $20^{circ}C$, whereas phytoplankton in ballast water treatment did not survive, regardless of optimal temperature. Colonization process was dominated by diatoms; Skeletonema coastatum for M/V Spring Lyra, Thalassiosira pseudonana and Thalassiosira for M/V Han Yang, Thalassiosira pacifica and Odontella aurita for M/V Modern Express, and Chaetoceros pseudocurvisetus and Pseudo-nitzschia seriata for M/V Asian Legend. The successful establishment of non-native species was also related to nutrient richness. Our laboratory design can be applied as a practical tool to assess the survivability of invasive microphytoplankton introduced into local waters of Ulsan and Pyeongtaek.