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LLC-PK1-CL4 cell expressing 3x-Citrine-Myo1a-Tail Homology 1 (TH1). Single molecule membrane binding events were captured using total internal reflection fluorescence (TIRF) microscopy.  Image stack (12.5 sec, 20 frames per second) was converted to a maximal intensity projection and subjected to three dimensional intensity profiling in Elements (Nikon).  Peaks represent ‘hot spots’ for long-lived membrane binding events.