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. Author manuscript; available in PMC: 2011 Jul 15.
Published in final edited form as: Neuron. 2010 Jul 15;67(1):86–99. doi: 10.1016/j.neuron.2010.05.026

Figure 4. Spatially restricted distribution of polymerized actin visualized with PALM near synapses.

Figure 4

A. Two-color PALM image of actin-tdEos and GKAP-dronpa in a fixed neuron. Molecules localized with error less than 30 nm (Actin-tdEos, red) or 100 nm (GKAP-Dronpa, green) are shown mapped with a pixel size of 5 nm. Scale bar, 1 μm.

B. Closer view of synapses on a spine (top) and shaft (bottom), showing lines along which measurements were made in C. Scale bar, 500 nm.

C. Relative molecular density probability of actin-tdEos (green) and GKAP-Dronpa (red) along 75-nm wide lines (30 pixels) shown in B. Dotted lines highlight the width of the profile at a probability of 0.5 (top, 121 nm; bottom, 87 nm).

D. The local density of polymerized actin calculated as the number of tracked actin-mEos2 molecules within a 111 nm radius. Local density (green) was plotted at ¼ this scale (27.8 nm bins), and overlaid on the diffraction-limited image of PSD-95-cerulean (red). Scale bar, 5 μm.

E. The distribution of tracked actin-mEos2 molecules as a function of distance from the center of PSDs identified by localization of PSD-95-cerulean (n = 4 neurons, 148 PSDs).

F–H. Three synapses from D, revealing close association of polymerizing actin with the PSD. Scale bars, 500 nm.

I. Actin distribution at a shaft synapse showing actin (left) encircling the PSD (middle). Scale bar, 500 nm. Green color bar represents local density of polymerized actin in A–E.