Live Visualization of Calcified Bones in Zebrafish and Medaka Larvae and Juveniles Using Calcein and Alizarin Red S
Zebrafish and medaka are valuable model vertebrates for genetic studies. The advent of CRISPR-Cas9 technology has greatly enhanced our capability to produce specific gene mutants in zebrafish and medaka. Analyzing the phenotypes of these mutants is essential for elucidating gene function, though suc...
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Bio-protocol LLC
2024-12-01
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| author | Rina Koita Sae Oikawa Taisei Tani Masaru Matsuda Akinori Kawamura |
| author_facet | Rina Koita Sae Oikawa Taisei Tani Masaru Matsuda Akinori Kawamura |
| author_sort | Rina Koita |
| collection | DOAJ |
| description | Zebrafish and medaka are valuable model vertebrates for genetic studies. The advent of CRISPR-Cas9 technology has greatly enhanced our capability to produce specific gene mutants in zebrafish and medaka. Analyzing the phenotypes of these mutants is essential for elucidating gene function, though such analyses often yield unexpected results. Consequently, providing researchers with accessible and cost-effective phenotype analysis methods is crucial. A prevalent technique for investigating calcified bone development in these species involves using transgenic fish that express fluorescent proteins labeling calcified bones; however, acquiring these fish and isolating appropriate crosses can be time-consuming. We present a comprehensive protocol for visualizing ossified bones in zebrafish and medaka larvae and juveniles using calcein and alizarin red S staining, which is both economical and efficient. This method, applicable to live specimens during the ossification of bones, avoids apparent alterations in skeletal morphology and allows for the use of different fluorescent dyes in conjunction with transgenic labeling, thus enhancing the analysis of developmental processes in calcifying bones, such as vertebrae and fin rays. |
| format | Article |
| id | doaj-art-1be403ef937b47859d2fa713c3eff4df |
| institution | Kabale University |
| issn | 2331-8325 |
| language | English |
| publishDate | 2024-12-01 |
| publisher | Bio-protocol LLC |
| record_format | Article |
| series | Bio-Protocol |
| spelling | doaj-art-1be403ef937b47859d2fa713c3eff4df2025-02-07T08:16:22ZengBio-protocol LLCBio-Protocol2331-83252024-12-01142410.21769/BioProtoc.5142Live Visualization of Calcified Bones in Zebrafish and Medaka Larvae and Juveniles Using Calcein and Alizarin Red SRina Koita0Sae Oikawa1Taisei Tani2Masaru Matsuda3Akinori Kawamura4Division of Life Science, Graduate School of Science and Engineering, Saitama University, Shimo-Okubo 255, Sakura-ku, Saitama, JapanCenter for Bioscience Research and Education, Utsunomiya University, 350 Mine-machi, Utsunomiya, JapanDivision of Life Science, Graduate School of Science and Engineering, Saitama University, Shimo-Okubo 255, Sakura-ku, Saitama, JapanDivision of Life Science, Graduate School of Science and Engineering, Saitama University, Shimo-Okubo 255, Sakura-ku, Saitama, JapanCenter for Bioscience Research and Education, Utsunomiya University, 350 Mine-machi, Utsunomiya, JapanDivision of Life Science, Graduate School of Science and Engineering, Saitama University, Shimo-Okubo 255, Sakura-ku, Saitama, JapanZebrafish and medaka are valuable model vertebrates for genetic studies. The advent of CRISPR-Cas9 technology has greatly enhanced our capability to produce specific gene mutants in zebrafish and medaka. Analyzing the phenotypes of these mutants is essential for elucidating gene function, though such analyses often yield unexpected results. Consequently, providing researchers with accessible and cost-effective phenotype analysis methods is crucial. A prevalent technique for investigating calcified bone development in these species involves using transgenic fish that express fluorescent proteins labeling calcified bones; however, acquiring these fish and isolating appropriate crosses can be time-consuming. We present a comprehensive protocol for visualizing ossified bones in zebrafish and medaka larvae and juveniles using calcein and alizarin red S staining, which is both economical and efficient. This method, applicable to live specimens during the ossification of bones, avoids apparent alterations in skeletal morphology and allows for the use of different fluorescent dyes in conjunction with transgenic labeling, thus enhancing the analysis of developmental processes in calcifying bones, such as vertebrae and fin rays.https://bio-protocol.org/en/bpdetail?id=5142&type=0 |
| spellingShingle | Rina Koita Sae Oikawa Taisei Tani Masaru Matsuda Akinori Kawamura Live Visualization of Calcified Bones in Zebrafish and Medaka Larvae and Juveniles Using Calcein and Alizarin Red S Bio-Protocol |
| title | Live Visualization of Calcified Bones in Zebrafish and Medaka Larvae and Juveniles Using Calcein and Alizarin Red S |
| title_full | Live Visualization of Calcified Bones in Zebrafish and Medaka Larvae and Juveniles Using Calcein and Alizarin Red S |
| title_fullStr | Live Visualization of Calcified Bones in Zebrafish and Medaka Larvae and Juveniles Using Calcein and Alizarin Red S |
| title_full_unstemmed | Live Visualization of Calcified Bones in Zebrafish and Medaka Larvae and Juveniles Using Calcein and Alizarin Red S |
| title_short | Live Visualization of Calcified Bones in Zebrafish and Medaka Larvae and Juveniles Using Calcein and Alizarin Red S |
| title_sort | live visualization of calcified bones in zebrafish and medaka larvae and juveniles using calcein and alizarin red s |
| url | https://bio-protocol.org/en/bpdetail?id=5142&type=0 |
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