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Pioneered &gt;30 years ago, IOSI allows macroscopic mapping of neuronal activity throughout the cortex. The technique has been used to study sensory processing and experience-dependent plasticity, and is often used as an adjunctive procedure to localize cortical areas for subsequent targeting by other imaging or physiology techniques. Despite the ubiquity of IOSI in neuroscience, there are few commercially available turn-key IOSI systems. As a result, investigators have typically resorted to building their own imaging systems. Over the years, simplified systems built either as dedicated rigs or incorporated into existing microscope platforms have been developed. Here we present a straightforward set of adaptations that can be applied to any standard upright microscope, using readily available, inexpensive, commercial parts for illumination, optics, and signal detection, that enables high-sensitivity IOSI. Using these adaptations, we are able to readily map sensory-evoked signals across the somatosensory and visual cortex, including single-whisker barrel cortical activity maps in mice. We show that these IOSI maps are highly reproducible across animals and can be used to study plasticity mechanisms in the somatosensory cortex. We also provide open-source applications to control illumination and analyze raw data to generate activity maps. We anticipate that these resources will be useful for neuroscience investigators looking to add IOSI capabilities to an existing microscope in the laboratory on a budget.<\/jats:p>","DOI":"10.1523\/eneuro.0046-23.2023","type":"journal-article","created":{"date-parts":[[2023,8,7]],"date-time":"2023-08-07T17:50:13Z","timestamp":1691430613000},"page":"ENEURO.0046-23.2023","source":"Crossref","is-referenced-by-count":15,"title":["High-Sensitivity Intrinsic Optical Signal Imaging Through Flexible, Low-Cost Adaptations of an Upright Microscope"],"prefix":"10.1523","volume":"10","author":[{"given":"Brenda","family":"Vasquez","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Baruc","family":"Campos","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ashley","family":"Cao","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Aye Theint","family":"Theint","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0567-2239","authenticated-orcid":false,"given":"William","family":"Zeiger","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"393","published-online":{"date-parts":[[2023,8,7]]},"reference":[{"key":"2023080710500814000_10.8.ENEURO.0046-23.2023.1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0109116"},{"key":"2023080710500814000_10.8.ENEURO.0046-23.2023.2","doi-asserted-by":"crossref","first-page":"100779","DOI":"10.1016\/j.xpro.2021.100779","article-title":"Intrinsic optical signal imaging and targeted injections through a chronic cranial window of a head-fixed mouse.","volume":"2","year":"2021","journal-title":"STAR Protoc"},{"key":"2023080710500814000_10.8.ENEURO.0046-23.2023.3","doi-asserted-by":"publisher","DOI":"10.1152\/jn.00729.2001"},{"key":"2023080710500814000_10.8.ENEURO.0046-23.2023.4","doi-asserted-by":"publisher","DOI":"10.1093\/cercor\/6.2.120"},{"key":"2023080710500814000_10.8.ENEURO.0046-23.2023.5","doi-asserted-by":"publisher","DOI":"10.1364\/OE.17.015670"},{"key":"2023080710500814000_10.8.ENEURO.0046-23.2023.6","doi-asserted-by":"crossref","first-page":"312","DOI":"10.1212\/WNL.55.2.312","article-title":"Optical intrinsic signal imaging in a rodent seizure model.","volume":"55","year":"2000","journal-title":"Neurology"},{"key":"2023080710500814000_10.8.ENEURO.0046-23.2023.7","doi-asserted-by":"publisher","DOI":"10.1038\/jcbfm.2013.7"},{"key":"2023080710500814000_10.8.ENEURO.0046-23.2023.8","doi-asserted-by":"publisher","DOI":"10.1016\/S0896-6273(00)80941-1"},{"key":"2023080710500814000_10.8.ENEURO.0046-23.2023.9","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2018.06.035"},{"key":"2023080710500814000_10.8.ENEURO.0046-23.2023.10","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0205444"},{"key":"2023080710500814000_10.8.ENEURO.0046-23.2023.11","doi-asserted-by":"publisher","DOI":"10.1101\/359513"},{"key":"2023080710500814000_10.8.ENEURO.0046-23.2023.12","doi-asserted-by":"publisher","DOI":"10.1093\/cercor\/bhn085"},{"key":"2023080710500814000_10.8.ENEURO.0046-23.2023.13","doi-asserted-by":"publisher","DOI":"10.1080\/08990220500084990"},{"key":"2023080710500814000_10.8.ENEURO.0046-23.2023.14","doi-asserted-by":"crossref","unstructured":"Frostig RD , Chen-Bee CH (2009) Visualizing adult cortical plasticity using intrinsic signal optical imaging. 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