biostudies-literatureUnknown4(1)Giacomoni JTissue clearing is commonly used for whole-brain imaging but seldom used for brain slices. Here, we present a simple protocol to slice, immunostain, and clear sections of adult rat brains for subsequent high-resolution confocal imaging. The protocol does not require toxic reagents or specialized equipment. We also provide instructions for culturing of rat brain slices free floating on permeable culture inserts, maintained in regular CO₂ incubators, and handled only at media change.STAR protocols102041https://www.ebi.ac.uk/biostudies/studies/S-EPMC9873499biostudies-literatureProtocol for optical clearing and imaging of fluorescently labeled ex vivo rat brain slices.PMC9873499Giacomoni JCepeda-Prado EMattsson BKajtez JOttosson DRHabekost MParmar MfalseProtocol for optical clearing and imaging of fluorescently labeled ex vivo rat brain slices.Tissue clearing is commonly used for whole-brain imaging but seldom used for brain slices. Here, we present a simple protocol to slice, immunostain, and clear sections of adult rat brains for subsequent high-resolution confocal imaging. The protocol does not require toxic reagents or specialized equipment. We also provide instructions for culturing of rat brain slices free floating on permeable culture inserts, maintained in regular CO₂ incubators, and handled only at media change.2023-01-01T00:00:00Z2023 Jan2025-04-22T02:26:59.171Z2025-04-05T20:19:40.326ZS-EPMC98734993685366810.1016/j.xpro.2022.102041