An optimized iDISCO+ protocol for tissue clearing and 3D analysis of oxytocin and vasopressin cell network in the...

Here, we present an optimized iDISCO+ protocol combining tissue clearing and light sheet microscopy to map the postnatal development of oxytocin and vasopressin neurons in mouse hypothalamus. We describe tissue preparation, immunostaining, clearing, and imaging. We then detail how to process the 3D cell dataset to analyze cell network using a point-based recording procedure that accurately maps neurons in the Allen brain atlas. This protocol can be applied to any neuronal population, in different brain regions and at different developmental stages. For complete details on the use and execution of this protocol, please refer to Soumier et al. (2021).¹.