Lung adenocarcinoma is the most common type of lung cancer. A close monitor of in vivo tumor development may help to better understand the pathogenesis and pathological processes of this disease. A bimodal imaging strategy has been developed, which is a very important tool to investigate the growth and metastasis of lung adenocarcinoma. In the present study, we used a combined labeling strategy in p53RE-luc-A549 cells via transfecting the reporter gene EGFP. In order to unambiguously identify the growth and metastasis of transfected A549 tumor cells, we established and observed subcutaneous and orthotopic xenografts in nude mice by in vivo bioluminescence and fluorescence imaging, which was verified by our post-mortem histological analysis. In vivo bioluminescence signal was observed for the progression of both subcutaneous and orthotopic xenografts in EGFP-p53RE-luc-A549 cells; in vivo fluorescence was only observed for the growth of subcutaneous xenograft of EGFP-p53RE-luc-A549 cells. Moreover, EGFP-p53RE-luc-A549 cells allow for the improved identification of implanted cells within host tissue during histological analysis. In conclusion, we presented a combined labeling strategy for bimodal A549 cell imaging which leads to improved detection of cellular grafts.