A single electron has been observed on a nanodot in a double-barrier tunneling structure by noncontact atomic-force microscopy at fixed separation. Frequency shift-voltage dependence of an Au-coated cantilever/vacuum/1-decanethiol protected Au nanodot/1-octanethiol self-assembled monolayer/Au substrate structure deviates from the theoretical parabolic curve, which is attributed to the change in the number of quantized electrons on the Au nanodot caused by the Coulomb blockade phenomena.