File:MacGillavry Blanpied 2013 Fig1.png

Figure 1

Figure 1. Superresolution Imaging of the Postsynaptic Density (A) Oblique illumination of SEP-GluA1 marking synapses along a stretch of dendrite, 100 nm pixels. (B) Same region, PALM image of shrPSD-95-mEos2, 25 nm pixels. Scale bar represents 500 nm. (C) Single-molecule localizations of PSD-95-PAtagRFP PALM (red) superimposed upon the widefield image of SEP-GluA1 fluorescence (white). Scale bars represent 1 mm (left) and 500 nm (right). (D and E) Histograms of number of detected photons per molecule (D) and localization precision (E). (F) Histogram of effective resolution calculated from the localization precision and density of shrPSD-95-mEos2 molecules in individual PSDs. (G) Cumulative distribution of PSD area measured by PALM for mEos2-tagged shrPSD-95 and shrShank3 compared with confocal microscopy of neurons expressing shrPSD-95-GFP. (H) Boxplot summary of PSD area measured by PALM for shrPSD-95 and shrShank3, compared with confocal, mean PSD area based on published EM data (see Results) and PALM on neurons overexpressing GKAP, Shank3, or Homer1c. **p < 0.01, Kruskal-Wallis. (I) Comparison of summed oblique illumination (top) and PALM image (bottom) of shrPSD-95-mEos2. Arrowheads mark PSDs expanded at right. Pixel size is 25 nm (left) and 12 nm (right). Scale bar represents 2.5 mm (left) and 200 nm (right).