An optimized and detailed step-by-step protocol for the analysis of neuronal morphology in golgi-stained fetal sheep brain
We describe the step-by-step protocol to retrieve neuronal morphometrics using Imaris imaging software to provide quantification of apical and basal dendrites for measures of dendrite length ( μm), branch number, branch order and Sholl analysis (intersections over radius). We also detail software add-ons for data retrieval of dendritic spines including the number of spines, spine density and spine classification, which are critical indicators of synaptic function. The assessment of neuro nal morphology in the developing brain using Rapid-Golgi and Imaris software is labour-intensive, particularly during the optimization period. The methodology described in this step-by-step description is novel, detailed, and aims to provide a reproducible, working protocol to quantify neuronal cyto architecture with simple descriptions that will save time for the next users of these commonly used techniques.