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Early detection of liver cancer and its biomarkers remains significant challenge.In this work, we report on the detection of alpha-fetoprotein (AFP), a surveillance biomarker of live cancer, by using in-situ atomic force microscopy (AFM) at a single-molecule level.AFM size measurements confirmed the successful capture of single AFP by monoclonal antibody molecules on a one-to-one basis.This method is straightforward and the results provide the direct evidence for the single molecule detection of AFP in the early diagnosis of liver cancer.