We report the observation of individual steps taken by motor proteins in living cells by following movements of endocytic vesicles that contain quantum dots (QDs) with a fast camera. The brightness and photostability of quantum dots allow us to record motor displacement traces with 300 mu s time resolution and 1.5 nm spatial precision. We observed individual 8 nm steps in active transport toward both the microtubule plus- and minus-ends, the directions of kinesin and dynein movements, respectively. In addition, we clearly resolved abrupt 16 nm steps in the plus-end direction and often consecutive 16 nm and occasional 24 nm steps in minus-end directed movements. This work demonstrates the ability of the QD assay to probe the operation of motor proteins at the molecular level in living cells under physiological conditions.