Cell migration is an important physiological process, which is among others controlled by ion channel activity. Calcium-activated potassium channels (K_Ca3.1) are required for optimal cell migration. Previously, we identified single hK_Ca3.1 channel proteins in the plasma membrane by means of quantum dot (QD) labeling. Here, we tracked single channel proteins during migration in order to classify their dynamics in the plasma membrane of MDCK-F cells. Single hK_Ca3.1 channels were visualised with QD- or Alexa488-conjugated antibodies and tracked at the basal cell membrane using time lapse TIRF microscopy. Analysis of the trajectories allowed the classification of channel dynamics. Channel tracks were compared with those of free QD-conjugated antibodies. The size of the label has a pronounced effect on hK_Ca3.1 channel diffusion. QD-labeled channels have a (sub-)diffusion coefficient D_QDbound = 0.067 µm²/s while that of Alexa488-labeled channels is D_alexa = 0.139 µm²/s. Free QD-conjugated antibodies move much faster: D_QDfree = 2.163 µm²/s. Plotting the mean squared distances (msd) covered by hK_Ca3.1 channels as function of time points to the mode of diffusion. Alexa488-labeled channels diffuse normally while the QD-label renders hK_Ca3.1 channel diffusion anomalous. Free QD-labeled antibodies also diffuse anomalously. Hence, QDs slow down diffusion of hK_Ca3.1 channels and change the mode of diffusion. These results, referring to the role of label size and properties of the extracellular environment, suggest that the pericellular glycocalyx has an important impact on labels used for single molecule tracking. Thus, tracking fluorescent particles within the glycocalyx opens up a possibility to characterize the pericellular nanoenvironement.