A novel method is presented for in situ quantification of living cell adhesion forces using a homemade nanorobotic system provided with two independently actuated probes that form a dual-probe nanotweezer capable of pick- and-place manipulation of a single living cell in an aqueous environment. Compared with single-cell force spectroscopy (SCFS) based on traditional atomic force microscopy (AFM), cell immobilization via chemical trapping is unnecessary and the test cell can be efficiently released using the nanotweezer to significantly enhance production of the SCFS. Benefiting from the accurate force sensing capability of AFM, the nanotweezer allows reliable force measurement ranging from picoNewtons to microNewtons and is sufficiently sensitive to characterize short- and long-term adhesion of cell-cell and cell-substrate adhesions. Capabilities of the nanotweezer have been validated through experimental qualification of cell-substrate and cell-cell adhesion events of C2C12 cells (mouse myoblast adherent) with different contact times.