Figure 2

Automated tracking of dendritic protrusions in live neurons. 11 DIV hippocampal neurons were transfected with a vector expressing Lifeact-ruby, and 24 hrs later were imaged at 5 min intervals for 1 hr. (a) Individual protrusions were tracked across the time series manually and with our automated method. This histogram shows the percentages of stable, new, and pruned protrusions for both methods. (b) Images depict 12 DIV neurons expressing Lifeact-ruby (white; top) overlayed with automated 3D reconstructions (bottom). The dendrite shaft is red and each tracked protrusion is labeled 1 through 4 (shades of blue and green). Images from t = 0 and 60 min are shown, and the scale bar is 5 μm. (c) Head width and (d) protrusion length were plotted versus time for each protrusion; the labels 1 through 4 in the legend correspond to the labels 1 through 4 in panel (b).