Engineered nanoparticles have emerged as potentially revolutionary drug and gene delivery vectors. Using rod-shaped gold nanoparticles as a model, we studied for the first time the rotational dynamics of nanoparticle vectors on live cell membranes and its impact on the fate of these nanoparticle vectors. The rotational motions of gold nanorods with various surface modifiers were tracked continuously at 200 frames/s under a differential interference contrast microscope. We found that the rotational behaviors of gold nanorod vectors are strongly related to their surface charges. Specific surface functional groups and the availability of receptors on cell membranes also contribute to the rotational dynamics. The study of rotational Brownian motion of nanoparticles on cell membranes will lead to a better understanding of the mechanisms of drug delivery and provide guidance in designing surface modification strategies for drug delivery vectors under various circumstances.