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Multi-Level Approach for Comprehensive Enamel Phenotyping
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SYSNO ASEP 0567377 Document Type J - Journal Article R&D Document Type Journal Article Subsidiary J Ostatní články Title Multi-Level Approach for Comprehensive Enamel Phenotyping Author(s) Aranaz Novalichez, Goretti (UMG-J)
Špoutil, František (UMG-J)
Buková, Ivana (UMG-J)
Krejzová, I. (CZ)
Olsinova, M. (CZ)
Dalecka, M. (CZ)
Benda, A. (CZ)
Rozman, Jan (UMG-J)
Sedláček, Radislav (UMG-J) RID
Procházka, Jan (UMG-J) ORCIDArticle number e340 Source Title Current Protocols - ISSN 2691-1299
Roč. 2, č. 1 (2022)Number of pages 20 s. Language eng - English Country US - United States Keywords Enamel ; MicroCT ; mouse models ; SEM ; teeth phenotyping OECD category Genetics and heredity (medical genetics to be 3) R&D Projects EF16_013/0001775 GA MŠMT - Ministry of Education, Youth and Sports (MEYS) LM2015040 GA MŠMT - Ministry of Education, Youth and Sports (MEYS) LM2018126 GA MŠMT - Ministry of Education, Youth and Sports (MEYS) EF16_013/0001789 GA MŠMT - Ministry of Education, Youth and Sports (MEYS) ED1.1.00/02.0109 GA MŠMT - Ministry of Education, Youth and Sports (MEYS) GJ19-19025Y GA ČR - Czech Science Foundation (CSF) ED2.1.00/19.0395 GA MŠMT - Ministry of Education, Youth and Sports (MEYS) Method of publishing Limited access DOI 10.1002/cpz1.340 Annotation 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 µm for in vivo imaging to 1.5 µm for ex vivo 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-carbon ratio. However, the methods mentioned above only show the result?already 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?starting with the live animal being scanned harmlessly and quickly, followed by sacrifice and high-resolution MicroCT scans requiring no special sample preparation. The biggest advantage is that samples remain in perfect condition for SEM or STED microscopic analysis. ? 2022 Wiley Periodicals LLC. Basic Protocol 1: In vivo MicroCT scanning of mouse Basic Protocol 2: Ex vivo HR-MicroCT of the teeth Basic Protocol 3: SEM for teeth microstructure Basic Protocol 4: Stimulated emission depletion (STED) microscopy Workplace Institute of Molecular Genetics Contact Nikol Škňouřilová, nikol.sknourilova@img.cas.cz, Tel.: 241 063 217 Year of Publishing 2023 Electronic address https://doi.org/10.1002/cpz1.340
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