Confocal microscopy was used to capture images of dendrites from the CA1 region of the hippocampus, based on previously described methods (75, 78). A blinded experimenter performed all imaging. Images were captured with a Nikon (Tokyo, Japan) Ti2 C2 confocal microscope. The experimenter identified secondary dendrites from dye-impregnated neurons and captured 3D z stacks of those meeting the following criteria: (i) within 80-μm working distance of microscope, (ii) relatively parallel with the surface of the coronal section, (iii) no overlap with other branches, (iv) minimum of 50 μm from the soma, and (v) maximum of 110 μm from the soma. For each dendrite, z stacks were captured with a 60× oil immersion objective (NA, 1.40; Nikon Plan Apo) using the following parameters: z step, 0.1 μm; image size; 1024 by 512 pixels (0.04 μm × 0.04 μm × 0.1 μm); zoom, 4.8×;, line averaging, 4; and acquisition rate: 1 frame/s. Captured images were deconvolved using Huygens Deconvolution System (16.05, Scientific Volume Imaging, The Netherlands) and the following settings: GMLE; maximum iterations, 10; signal-to-noise ratio, 15; and quality, 0.003. Deconvolved images were saved in .tif format.

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