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While such procedures have been successful in mice, few reports provide detailed procedures for successful surgery and long\u2010term imaging in rats, which are better suited for studying complex human behaviors. We present a robotic surgical protocol for same\u2010day virus injection and GRIN lens implantation into the rat nucleus accumbens core. Our procedure utilizes a direct lens insertion without tissue aspiration and produces quality image retention for months of recording. We also describe daily recording strategies to minimize damage and promote long\u2010term imaging. Finally, we present custom protective strategies to eliminate the need to remove miniscopes between sessions. This methodology protects rats from repeated isoflurane exposure and ensures a consistent focal plane for the entirety of the experiment. \u00a9 2025 The Author(s). Current Protocols published by Wiley Periodicals LLC.<\/jats:p><jats:p><jats:bold>Basic Protocol 1<\/jats:bold>: Craniotomy<\/jats:p><jats:p><jats:bold>Basic Protocol 2<\/jats:bold>: Virus injection<\/jats:p><jats:p><jats:bold>Basic Protocol 3<\/jats:bold>: GRIN lens implantation<\/jats:p><jats:p><jats:bold>Basic Protocol 4<\/jats:bold>: Baseplate mounting and assessment of the anesthetized rat<\/jats:p><jats:p><jats:bold>Basic Protocol 5<\/jats:bold>: Assessment of the awake, behaving rat<\/jats:p><jats:p><jats:bold>Support Protocol 1<\/jats:bold>: Protective miniscope cone fabrication<\/jats:p><jats:p><jats:bold>Support Protocol 2<\/jats:bold>: Miniscope cable fabrication<\/jats:p>","DOI":"10.1002\/cpz1.70090","type":"journal-article","created":{"date-parts":[[2025,1,10]],"date-time":"2025-01-10T02:08:18Z","timestamp":1736474898000},"source":"Crossref","is-referenced-by-count":4,"title":["Miniscope Imaging of Nucleus Accumbens Neural Activity in Freely Behaving Rats: Virus Injection, Gradient Index Lens Implantation, Recording Strategies, and Analytical Methods"],"prefix":"10.1002","volume":"5","author":[{"given":"Nicholas J.","family":"Beacher","sequence":"first","affiliation":[{"name":"Intramural Research Program National Institute on Drug Abuse  Baltimore Maryland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0002-9595-4415","authenticated-orcid":false,"given":"Michael W.","family":"Wang","sequence":"additional","affiliation":[{"name":"Intramural Research Program National Institute on Drug Abuse  Baltimore Maryland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Matthew C.","family":"Broomer","sequence":"additional","affiliation":[{"name":"Intramural Research Program National Institute on Drug Abuse  Baltimore Maryland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jessica Y.","family":"Kuo","sequence":"additional","affiliation":[{"name":"University of California Davis Health, School of Medicine, Department of Neurology Alzheimer's Disease Research Center\u2013East Bay  Walnut Creek California"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kayden A.","family":"Washington","sequence":"additional","affiliation":[{"name":"Feinberg School of Medicine Northwestern University  Chicago Illinois"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Miranda","family":"Targum","sequence":"additional","affiliation":[{"name":"Penn Memory Center University of Pennsylvania  Philadelphia Pennsylvania"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yan","family":"Zhang","sequence":"additional","affiliation":[{"name":"National Institute on Drug Dependence and Beijing Key Laboratory of Drug Dependence Peking University  Beijing China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Giovanni","family":"Barbera","sequence":"additional","affiliation":[{"name":"Intramural Research Program National Institute on Drug Abuse  Baltimore Maryland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0538-7892","authenticated-orcid":false,"given":"Da\u2010Ting","family":"Lin","sequence":"additional","affiliation":[{"name":"Intramural Research Program National Institute on Drug Abuse  Baltimore Maryland"},{"name":"The Solomon H. 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