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Aim: To investigate the movement of α1A-adrenergic receptors(α1A-AR) stimu-lated by agonist, phenylephrine (PE), and the dynamics of receptor movement in real time in single living cells with millisecond resolution.Methods: We labeled α1A-AR using the monoclonal, anti-FLAG (a kind of tag) antibody and Cy3-conju-gated goat anti-mouse IgG and recorded the trajectory of their transport process in living HEK293A cells stimulated by agonist, PE, and then analyzed their dy-namic properties.Results: The specific detection of α1A-AR on the surface of living HEK293A-α1A cells was achieved. α1A-AR intealize under the stimulation of PE. After the cells were stimulated with PE for 20 rain, apparent colocalization was found between α1A-AR and F-actins. After 40 min stimulation of PE, trajecto-ries of approximate linear motion in HEK293A-α1A cells were recorded, and their velocity was calculated. Condusion: The specific labeling method on the living cell surface provides a convenient means of real-time detection of the behavior of surface receptors. By this method we were able to specifically detectα1A-AR and record the behavior of individual particles of receptors with 50 ms exposure time in real time in single living cells.