Objective To determine the target recognition ability of the near-infrared fluorescence (NIRF) heptamethine cyanine dye in mouse models of orthotopically transplanted gastric carcinoma with optical imaging. Methods The orthotopically transplanted model of gastric carcinoma was established by implantation of luciferase-tagged-HepG2 cells into the stomach in nude mice, and gastric ulcer model was induced by absolute ethanol. Both bioluminescence (BIL) signal and NIRF signal in those two animal models were observed with optical imaging respectively, and the absorption of NIRF dye in gastric carcinoma tissues was determined. We further explored the effect of hypoxia and OATP on the absorption of the NIRF dye in gastric carcinoma tissues. The specific targeting ability of NIRF dye to tumor cells was evaluated. Result A good positive correlation was observed between NIRF signal and BIL signal (R²=0.995). Strong NIRF signal was observed in gastric carcinoma region, but no signal was found in the gastric ulcer model. Moreover, hypoxia further promoted the uptake of NIRF dye in gastric carcinoma, but OATP specific inhibitor BSP significantly reduced the absorption of NIRF dye in tumor cells. Conclusions The NIRF heptamethine cyanine dye can be applied to identify the orthotopically transplanted gastric carcinoma in nude mouse models.