Tumor marker detection contributes to the early diagnosis of cancers. However, due to the lack of detection specificity, its results cannot act as a direct evidence to confirm cancer occurrence in clinic. Joint detection of tumor markers may improve the detection specificity. As a trial for clinical diagnosis of hepatocellular carcinoma, alpha-fetoprotein, alpha-L-fucosidase and ferritin have been combined and detected with a label-free, phase sensitive and high throughput imaging ellipsometry biosensor (IEB). Eighty-two sera have been quantitatively detected with IEB and the results are in agreement with the clinical standard approaches. Evaluated by receiver operating characteristic analysis, the specificity of joint detection improves remarkably with IEB for hepatocellular carcinoma. It can be foreseen that the joint detection of tumor markers with IEB has a potential for clinical cancer diagnosis. (C) 2014 Elsevier B.V. All rights reserved.