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. Author manuscript; available in PMC: 2012 Jul 19.
Published in final edited form as: Nat Methods. 2011 Sep 18;8(11):969–975. doi: 10.1038/nmeth.1704

Figure 1.

Figure 1

Single molecules appear multiple times with variable blinking intervals. (a–c) TfR-PAGFP was expressed transiently at low or high density on the plasma membrane of COS-7 cells and imaged using PALM. Best fit coordinates of peak centers from individual frames were calculated and displayed as single dots in a and c. Shown are clusters of peaks corresponding to two spatially and temporally well-separated molecules in low expressing cell (a) and peaks corresponding to molecules that are not well separated in highly expressing cell (c). Note that the peaks arising from a single molecule are spatially confined in a region determined by average localization precision of the peaks. Center positions of molecules (asterisks) were determined by grouping peaks in the clusters. Color bars indicate frame number at which the molecules appeared. Graphs in b show multiple frame appearances of the single molecules (i) and (ii) from a. Frames in the sequence with no signal represent times when the molecule switched to a dark state.