{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,7,30]],"date-time":"2025-07-30T09:45:18Z","timestamp":1753868718194,"version":"3.41.2"},"reference-count":17,"publisher":"Wiley","issue":"1","license":[{"start":{"date-parts":[[2022,1,10]],"date-time":"2022-01-10T00:00:00Z","timestamp":1641772800000},"content-version":"vor","delay-in-days":9,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Current Protocols"],"published-print":{"date-parts":[[2022,1]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Enamel is the hardest tissue in mammalian organisms and is the layer covering the tooth. It consists of hydroxyapatite (HAP) crystallites, which mineralize on a protein scaffold known as the enamel matrix. Enamel matrix assembly is a very complex process mediated by enamel matrix proteins (EMPs). Altered HAP deposition or disintegration of the protein scaffold can cause enamel defects. Various methods have been established for enamel phenotyping, including MicroCT scanning with various resolutions from 9 \u00b5m for <jats:italic>in vivo<\/jats:italic> imaging to 1.5 \u00b5m for <jats:italic>ex vivo<\/jats:italic> imaging. With increasing resolution, we can see not only the enamel layer itself but also a detailed map of mineralization. To study enamel microstructure, we combine the MicroCT analysis with scanning electron microscopy (SEM), which enables us to perform element analyses such as calcium\u2010carbon ratio. However, the methods mentioned above only show the result\u2014already formed enamel. Stimulated emission depletion (STED) microscopy provides extra information about protein structure in the form of EMP localization and position before enamel mineralization. A combination of all these methods allows analyzing the same sample on multiple levels\u2014starting with the live animal being scanned harmlessly and quickly, followed by sacrifice and high\u2010resolution MicroCT scans requiring no special sample preparation. The biggest advantage is that samples remain in perfect condition for SEM or STED microscopic analysis. \u00a9 2022 Wiley Periodicals LLC.<\/jats:p><jats:p><jats:bold>Basic Protocol 1<\/jats:bold>: In vivo MicroCT scanning of mouse<\/jats:p><jats:p><jats:bold>Basic Protocol 2<\/jats:bold>: <jats:italic>Ex vivo<\/jats:italic> HR\u2010MicroCT of the teeth<\/jats:p><jats:p><jats:bold>Basic Protocol 3<\/jats:bold>: SEM for teeth microstructure<\/jats:p><jats:p><jats:bold>Basic Protocol 4<\/jats:bold>: Stimulated emission depletion (STED) microscopy<\/jats:p>","DOI":"10.1002\/cpz1.340","type":"journal-article","created":{"date-parts":[[2022,1,10]],"date-time":"2022-01-10T20:39:26Z","timestamp":1641847166000},"source":"Crossref","is-referenced-by-count":0,"title":["Multi\u2010Level Approach for Comprehensive Enamel Phenotyping"],"prefix":"10.1002","volume":"2","author":[{"given":"Goretti","family":"Aranaz\u2010Novaliches","sequence":"first","affiliation":[{"name":"Laboratory of Transgenic Models of Diseases Institute of Molecular Genetics of the Czech Academy of Sciences  Prague Czech Republic"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Frantisek","family":"Spoutil","sequence":"additional","affiliation":[{"name":"Czech Centre for Phenogenomics Institute of Molecular Genetics of the Czech Academy of Sciences  Prague Czech Republic"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ivana","family":"Bukova","sequence":"additional","affiliation":[{"name":"Laboratory of Transgenic Models of Diseases Institute of Molecular Genetics of the Czech Academy of Sciences  Prague Czech Republic"},{"name":"Czech Centre for Phenogenomics Institute of Molecular Genetics of the Czech Academy of Sciences  Prague Czech Republic"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Irena","family":"Krejzova","sequence":"additional","affiliation":[{"name":"Imaging Methods Core Facility Charles University in Prague, Faculty of Science  Prague Czech Republic"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Marie","family":"Olsinova","sequence":"additional","affiliation":[{"name":"Imaging Methods Core Facility Charles University in Prague, Faculty of Science  Prague Czech Republic"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Marketa","family":"Dalecka","sequence":"additional","affiliation":[{"name":"Imaging Methods Core Facility Charles University in Prague, Faculty of Science  Prague Czech Republic"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ales","family":"Benda","sequence":"additional","affiliation":[{"name":"Imaging Methods Core Facility Charles University in Prague, Faculty of Science  Prague Czech Republic"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jan","family":"Rozman","sequence":"additional","affiliation":[{"name":"Czech Centre for Phenogenomics Institute of Molecular Genetics of the Czech Academy of Sciences  Prague Czech Republic"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Radislav","family":"Sedlacek","sequence":"additional","affiliation":[{"name":"Laboratory of Transgenic Models of Diseases Institute of Molecular Genetics of the Czech Academy of Sciences  Prague Czech Republic"},{"name":"Czech Centre for Phenogenomics Institute of Molecular Genetics of the Czech Academy of Sciences  Prague Czech Republic"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jan","family":"Prochazka","sequence":"additional","affiliation":[{"name":"Laboratory of Transgenic Models of Diseases Institute of Molecular Genetics of the Czech Academy of Sciences  Prague Czech Republic"},{"name":"Czech Centre for Phenogenomics Institute of Molecular Genetics of the Czech Academy of Sciences  Prague Czech Republic"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2022,1,10]]},"reference":[{"key":"e_1_2_8_2_1","doi-asserted-by":"publisher","DOI":"10.1007\/978-3-642-83496-7_6"},{"key":"e_1_2_8_3_1","doi-asserted-by":"publisher","DOI":"10.1177\/154405910708600405"},{"key":"e_1_2_8_4_1","doi-asserted-by":"publisher","DOI":"10.1002\/0471142735.im0106s73"},{"key":"e_1_2_8_5_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41598\u2010021\u201094303\u20104"},{"key":"e_1_2_8_6_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M104624200"},{"key":"e_1_2_8_7_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0089303"},{"key":"e_1_2_8_8_1","doi-asserted-by":"publisher","DOI":"10.7860\/JCDR\/2015\/14641.6569"},{"key":"e_1_2_8_9_1","doi-asserted-by":"publisher","DOI":"10.1152\/physrev.00030.2016"},{"key":"e_1_2_8_10_1","doi-asserted-by":"publisher","DOI":"10.1177\/0022034516645389"},{"key":"e_1_2_8_11_1","first-page":"325","article-title":"Morphological characteristics of mouse incisor enamel","volume":"189","author":"M\u00f8inichen C. 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