A rapid and nondestructive protocol for whole-mount bone staining of small fish and Xenopus
Here we propose a new protocol for whole-mount bone staining, which allows the rapid preparation of highly cleared and nondestructive specimens. It only takes 3 days to complete whole procedure for small vertebrates, such as medaka, zebrafish, and Xenopus frogs. In this procedure, we used a newly de...
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| Veröffentlicht in: | Scientific reports 2018-05, Vol.8 (1), p.7453-7, Article 7453 |
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| creator | Sakata-Haga, Hiromi Uchishiba, Maimi Shimada, Hiroki Tsukada, Tsuyoshi Mitani, Mayumi Arikawa, Tomohiro Shoji, Hiroki Hatta, Toshihisa |
| description | Here we propose a new protocol for whole-mount bone staining, which allows the rapid preparation of highly cleared and nondestructive specimens. It only takes 3 days to complete whole procedure for small vertebrates, such as medaka, zebrafish, and
Xenopus
frogs. In this procedure, we used a newly developed fixative containing formalin, Triton X-100, and potassium hydroxide, which allows the fixation, decolorization, and transparentization of specimens at the same time. A bone staining solution containing alizarin red S with ethylene glycol and a clearing solution containing Tween 20 and potassium hydroxide also contributed the specificity and swiftness of this new system. As expected, although details of the skeletal system could be observed in specimens with high transparency, it was noteworthy that high-resolution fluorescence images acquired using zoom microscopes clearly delineated the shape of each bone. This new procedure would be expected to be widely used as a standard procedure for bone staining in the testing the developmental toxicity of chemicals and in the screening test of knockout or mutant animals. |
| doi_str_mv | 10.1038/s41598-018-25836-4 |
| format | Article |
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Xenopus
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Uchishiba, Maimi ; Shimada, Hiroki ; Tsukada, Tsuyoshi ; Mitani, Mayumi ; Arikawa, Tomohiro ; Shoji, Hiroki ; Hatta, Toshihisa</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c540t-a7a297c408347b5aae8e81b92672ec3fbd37d13f08ee1f26532775f95fd32d5e3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2018</creationdate><topic>14/34</topic><topic>14/63</topic><topic>631/136/818</topic><topic>631/154/570</topic><topic>64/114</topic><topic>64/116</topic><topic>Decolorization</topic><topic>Ethylene glycol</topic><topic>Fluorescence in situ hybridization</topic><topic>Humanities and Social Sciences</topic><topic>Microscopes</topic><topic>multidisciplinary</topic><topic>Potash</topic><topic>Potassium</topic><topic>Potassium hydroxide</topic><topic>Science</topic><topic>Science (multidisciplinary)</topic><topic>Skeletal system</topic><topic>Toxicity</topic><topic>Xenopus</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Sakata-Haga, Hiromi</creatorcontrib><creatorcontrib>Uchishiba, Maimi</creatorcontrib><creatorcontrib>Shimada, Hiroki</creatorcontrib><creatorcontrib>Tsukada, Tsuyoshi</creatorcontrib><creatorcontrib>Mitani, Mayumi</creatorcontrib><creatorcontrib>Arikawa, Tomohiro</creatorcontrib><creatorcontrib>Shoji, Hiroki</creatorcontrib><creatorcontrib>Hatta, Toshihisa</creatorcontrib><collection>Springer Nature OA Free Journals</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>ProQuest Central (Corporate)</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Biology Database (Alumni Edition)</collection><collection>Medical Database (Alumni Edition)</collection><collection>Science Database (Alumni Edition)</collection><collection>ProQuest SciTech Collection</collection><collection>ProQuest Natural Science Collection</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>ProQuest Central (Alumni Edition)</collection><collection>ProQuest One Sustainability</collection><collection>ProQuest Central UK/Ireland</collection><collection>ProQuest Central Essentials</collection><collection>Biological Science Collection</collection><collection>ProQuest Central</collection><collection>Natural Science Collection</collection><collection>ProQuest One Community College</collection><collection>ProQuest Central Korea</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Central Student</collection><collection>SciTech Premium Collection</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>ProQuest Biological Science Collection</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>Medical Database</collection><collection>Science Database</collection><collection>Biological Science Database</collection><collection>Publicly Available Content Database</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>ProQuest Central Basic</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Scientific reports</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Sakata-Haga, Hiromi</au><au>Uchishiba, Maimi</au><au>Shimada, Hiroki</au><au>Tsukada, Tsuyoshi</au><au>Mitani, Mayumi</au><au>Arikawa, Tomohiro</au><au>Shoji, Hiroki</au><au>Hatta, Toshihisa</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>A rapid and nondestructive protocol for whole-mount bone staining of small fish and Xenopus</atitle><jtitle>Scientific reports</jtitle><stitle>Sci Rep</stitle><addtitle>Sci Rep</addtitle><date>2018-05-10</date><risdate>2018</risdate><volume>8</volume><issue>1</issue><spage>7453</spage><epage>7</epage><pages>7453-7</pages><artnum>7453</artnum><issn>2045-2322</issn><eissn>2045-2322</eissn><abstract>Here we propose a new protocol for whole-mount bone staining, which allows the rapid preparation of highly cleared and nondestructive specimens. It only takes 3 days to complete whole procedure for small vertebrates, such as medaka, zebrafish, and
Xenopus
frogs. In this procedure, we used a newly developed fixative containing formalin, Triton X-100, and potassium hydroxide, which allows the fixation, decolorization, and transparentization of specimens at the same time. A bone staining solution containing alizarin red S with ethylene glycol and a clearing solution containing Tween 20 and potassium hydroxide also contributed the specificity and swiftness of this new system. As expected, although details of the skeletal system could be observed in specimens with high transparency, it was noteworthy that high-resolution fluorescence images acquired using zoom microscopes clearly delineated the shape of each bone. This new procedure would be expected to be widely used as a standard procedure for bone staining in the testing the developmental toxicity of chemicals and in the screening test of knockout or mutant animals.</abstract><cop>London</cop><pub>Nature Publishing Group UK</pub><pmid>29748567</pmid><doi>10.1038/s41598-018-25836-4</doi><tpages>7</tpages><oa>free_for_read</oa></addata></record> |
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| subjects | 14/34 14/63 631/136/818 631/154/570 64/114 64/116 Decolorization Ethylene glycol Fluorescence in situ hybridization Humanities and Social Sciences Microscopes multidisciplinary Potash Potassium Potassium hydroxide Science Science (multidisciplinary) Skeletal system Toxicity Xenopus |
| title | A rapid and nondestructive protocol for whole-mount bone staining of small fish and Xenopus |
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