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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 2.

Figure 2

PC-PALM distinguishes random versus clustered distributions. (a) Diagram showing purified PAGFP molecules randomly immobilized on glass coverslip. (b) Actual spatial distribution of peak centers of PAGFP molecules in a section of this coverslip. (c) Plot of calculated autocorrelation function (g(r)peaks) of PAGFP molecules in b fit to equation (1). The correlation owing to multiple appearances of a single protein (g(r)stoch) and the protein correlation (g(r)protein) were evaluated from the fit. (d) Distribution of peak centers of TfR-PAGFP across the plasma membrane of a COS-7 cell. A representative section of the cell used for correlation analysis is marked by a yellow box. (e) Spatial distribution of peak centers of TfR-PAGFP in the section indicated in d. Beside peak clusters associated with individual molecules, there was noticeable grouping of these clusters (red dashed ovals). (f) Measured correlation function of all peaks (g(r)peaks) in e was well fit to equation (2). Corrected protein correlation function (g(r)protein) was evaluated by subtracting the contribution from multiple appearances of single protein (g(r)stoch) from the measured correlation function. Scale bars, 200 nm (b,e) and 5 μm (d).