{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,18]],"date-time":"2026-07-18T05:30:43Z","timestamp":1784352643342,"version":"3.55.0"},"reference-count":46,"publisher":"MDPI AG","issue":"11","license":[{"start":{"date-parts":[[2023,6,1]],"date-time":"2023-06-01T00:00:00Z","timestamp":1685577600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Alberta Innovates"},{"name":"Multiple Sclerosis Society of Canada"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Molecules"],"abstract":"<jats:p>Amyloids are misfolded proteins that aggregate into fibrillar structures, the accumulation of which is associated with the pathogenesis of many neurodegenerative diseases, such as Alzheimer\u2019s disease (AD). Early, sensitive detection of these misfolded aggregates is of great interest to the field, as amyloid deposition begins well before the presentation of clinical symptoms. Thioflavin-S (ThS) is a fluorescent probe commonly used to detect amyloid pathology. Protocols for ThS staining vary, but they often use high staining concentrations followed by differentiation, which causes varying levels of non-specific staining and potentially leaves more subtle amyloid deposition unidentified. In this study, we developed an optimized ThS staining protocol for the sensitive detection of \u03b2-amyloids in the widely used 5xFAD Alzheimer\u2019s mouse model. Controlled dye concentrations together with fluorescence spectroscopy and advanced analytical methods enabled not only the visualization of plaque pathology, but also the detection of subtle and widespread protein misfolding throughout the 5xFAD white matter and greater parenchyma. Together, these findings demonstrate the efficacy of a controlled ThS staining protocol and highlight the potential use of ThS for the detection of protein misfolding that precedes clinical manifestation of disease.<\/jats:p>","DOI":"10.3390\/molecules28114483","type":"journal-article","created":{"date-parts":[[2023,6,1]],"date-time":"2023-06-01T04:21:27Z","timestamp":1685593287000},"page":"4483","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":17,"title":["Quantitation of Tissue Amyloid via Fluorescence Spectroscopy Using Controlled Concentrations of Thioflavin-S"],"prefix":"10.3390","volume":"28","author":[{"ORCID":"https:\/\/orcid.org\/0009-0007-9324-3798","authenticated-orcid":false,"given":"Tatiana P.","family":"MacKeigan","sequence":"first","affiliation":[{"name":"Hotchkiss Brain Institute, Cumming School of Medicine, Department of Clinical Neurosciences, University of Calgary, 3330 Hospital Drive NW, Calgary, AB T2N 4N1, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2329-1413","authenticated-orcid":false,"given":"Megan L.","family":"Morgan","sequence":"additional","affiliation":[{"name":"Hotchkiss Brain Institute, Cumming School of Medicine, Department of Clinical Neurosciences, University of Calgary, 3330 Hospital Drive NW, Calgary, AB T2N 4N1, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0326-4194","authenticated-orcid":false,"given":"Peter K.","family":"Stys","sequence":"additional","affiliation":[{"name":"Hotchkiss Brain Institute, Cumming School of Medicine, Department of Clinical Neurosciences, University of Calgary, 3330 Hospital Drive NW, Calgary, AB T2N 4N1, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2023,6,1]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"237","DOI":"10.1007\/s12013-011-9200-x","article-title":"Defective Protein Folding and Aggregation as the Basis of Neurodegenerative Diseases: The Darker Aspect of Proteins","volume":"61","author":"Naeem","year":"2011","journal-title":"Cell Biochem. 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