Quantifying Neuronal Cell Populations

A VS120-S6-W Virtual Slide Scanner (Olympus) was used to scan all the sections. Images were taken with a color camera (Nikon DS-Fi3). To restrict any influences on our counted results, the photomicrography and counting were performed by one blind researcher. ImageJ (version 1.41; National Institutes of Health, Bethesda, MD) was used for cell counting and Canvas software (ACD Systems, Victoria, Canada, v. 9.0) was used for line drawings. A one-in-two series of 25 µm brain sections was used per mouse, which means that each section analyzed was 50 µm apart. The area analyzed was delimited based on previous reports (Anderson et al., 2016 (link)) (mean of 5423 μm²). The sections were counted bilaterally, averaged, and the numbers reported as mean ± standard error of the mean (SEM). Section alignment were relative to a reference section, as previously described (Anderson et al., 2016 (link)) and based on Paxinos and Franklin, 2019 .