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. 2021 Apr 5;2(2):100427. doi: 10.1016/j.xpro.2021.100427

Figure 2.

Figure 2

Comparison of pre- and post-deconvolution 3D dendritic reconstructions of spines and emerging spinules acquired using standard or optimized parameters

Three-dimensional IMARIS reconstruction confocal images depicting a secondary apical dendrite, mushroom spines, and emerging spinules from a DIV 21, GFP-expressing cortical pyramidal neuron.

(A) Live time-lapse confocal Z-stacks acquired using standard settings, i.e., Z-step=0.15 μm, pinhole radius=67.69, or settings optimized for enhanced resolution, i.e., Z-step=0.1 μm, pinhole radius=30.65, are shown before and after 3D automatic iterative deconvolution. Blue arrows and insets highlight the resolution of a fine, elongated, long-lived spinule protruding from a mushroom spine in each of the images. Scale bar equals 5 microns. See also Methods videos S1 and S2.

(B) Montage from Methods video S2 of a small mushroom spine forming a short-lived spinule (red arrow) that appears in a single time-point, with an estimated lifespan of ≤12 s. Scale bar equals 1 micron.

(C) Montage from Methods video S2 highlights a very long-lived spinule (blue arrows), with a lifespan equal to or exceeding the 15 min imaging duration, emerging from a large mushroom spine. Scale bar equals 1 micron.