{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T15:57:36Z","timestamp":1785427056831,"version":"3.56.0"},"posted":{"date-parts":[[2025,8,2]]},"group-title":"Bioengineering","reference-count":30,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2025,8,2]],"date-time":"2025-08-02T00:00:00Z","timestamp":1754092800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc\/4.0\/"}],"funder":[{"award":["2R15GM128166-02"],"award-info":[{"award-number":["2R15GM128166-02"]}],"id":[{"id":"https:\/\/ror.org\/01cwqze88","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2025,8,2]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>Spinning disk confocal microscopy enables fast optical sectioning with low phototoxicity but is often inaccessible due to high hardware costs. We present a low-cost solution using a 25 megapixel machine vision CMOS camera (Sony IMX540, FLIR Blackfly S) and a custom-built spinning disk. The system uses a back-illuminated sensor with high quantum efficiency (69% at 525 nm) and low read noise (2.31 electrons). High-resolution images of Thy1-GFP mouse brain slices and H&amp;E-stained rat testis verified performance across 3D tissue volumes. The custom disk, made with 18 \u00b5m pinholes (180 \u00b5m pitch) on a chrome photomask and mounted to an optical chopper motor, enables stable, near-telecentric imaging. Micro-Manager software integration allows synchronized control of all hardware, which demonstrates that affordable CMOS sensors can potentially replace sCMOS in spinning disk microscopy, offering an open-source, scalable solution for advanced imaging.<\/jats:p>","DOI":"10.1101\/2025.07.30.667773","type":"posted-content","created":{"date-parts":[[2025,8,2]],"date-time":"2025-08-02T16:45:11Z","timestamp":1754153111000},"source":"Crossref","is-referenced-by-count":0,"title":["Spinning disk confocal microscopy with a 25 Megapixel Camera"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-4802-9481","authenticated-orcid":false,"given":"Guy M.","family":"Hagen","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Brian","family":"Lewis","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Summer","family":"Levis","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Joseph R.","family":"Hamilton","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tristan C.","family":"Paul","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"key":"2025080518300462000_2025.07.30.667773v1.1","doi-asserted-by":"crossref","first-page":"1550","DOI":"10.1038\/s41467-024-45567-7","article-title":"Open-source microscope add-on for structured illumination microscopy","volume":"15","year":"2024","journal-title":"Nat. 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