{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,18]],"date-time":"2026-09-18T23:44:54Z","timestamp":1789775094312,"version":"4.0.1"},"reference-count":46,"publisher":"Public Library of Science (PLoS)","issue":"3","license":[{"start":{"date-parts":[[2025,3,31]],"date-time":"2025-03-31T00:00:00Z","timestamp":1743379200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100024872","name":"Applied and Engineering Sciences","doi-asserted-by":"crossref","award":["15825"],"award-info":[{"award-number":["15825"]}],"id":[{"id":"10.13039\/501100024872","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/501100003246","name":"Nederlandse Organisatie voor Wetenschappelijk Onderzoek","doi-asserted-by":"publisher","award":["1228.191.204"],"award-info":[{"award-number":["1228.191.204"]}],"id":[{"id":"10.13039\/501100003246","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100003000","name":"Stichting MS Research","doi-asserted-by":"publisher","award":["MS16-945b"],"award-info":[{"award-number":["MS16-945b"]}],"id":[{"id":"10.13039\/501100003000","id-type":"DOI","asserted-by":"publisher"}]},{"name":"National MS society","award":["PA2021-36033"],"award-info":[{"award-number":["PA2021-36033"]}]},{"name":"Dutch National MS Foundation","award":["OZ2021-008"],"award-info":[{"award-number":["OZ2021-008"]}]}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS ONE"],"abstract":"<jats:p>Myelin pathology is known to play a central role in disorders such as multiple sclerosis (MS) among others. Despite this, the pathological mechanisms underlying these conditions are often difficult to unravel. Conventional techniques like immunohistochemistry or dye-based approaches, do not provide a temporal characterization of the pathophysiological aberrations responsible for myelin changes in human specimens. Here, to circumvent this curb, we present a label-free, live-cell imaging approach of myelin using recent advancements in nonlinear harmonic generation microscopy applied to physiologically viable human brain tissue from post-mortem donors. Gray and white matter brain tissue from epilepsy surgery and post-mortem donors was excised. To sustain viability of the specimens for several hours, they were subjected to either acute or organotypic slice culture protocols in artificial cerebral spinal fluid. Imaging was performed using a femtosecond pulsed 1050\u2009nm laser to generate second harmonic generation (SHG) and third harmonic generation (THG) signals directly from myelin and axon-like structures without the need to add any labels. Experiments on acute human brain slices and post-mortem human slice cultures reveal that myelin, along with lipid bodies, are the prime sources of THG signal. We show that tissue viability is maintained over extended periods during THG microscopy, and that prolonged THG imaging is able to detect experimentally induced subtle alterations in myelin morphology. Finally, we provide practical evidence that live-cell imaging of myelin with THG microscopy is a sensitive tool to investigate subtle changes in white matter of neurological donors.<\/jats:p>\n                  <jats:p>Overall, our findings support that nonlinear live-cell imaging is a suitable setup for researching myelin morphology in neurological conditions like MS.<\/jats:p>","DOI":"10.1371\/journal.pone.0310663","type":"journal-article","created":{"date-parts":[[2025,3,31]],"date-time":"2025-03-31T16:23:02Z","timestamp":1743438182000},"page":"e0310663","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":9,"title":["Dynamic imaging of myelin pathology in physiologically preserved human brain tissue using third harmonic generation microscopy"],"prefix":"10.1371","volume":"20","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-2561-0206","authenticated-orcid":true,"given":"Niels R. 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