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A rapid and nondestructive protocol for whole-mount bone staining of small fish and Xenopus.

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.

SUBMITTER: Sakata-Haga H

PROVIDER: S-EPMC5945591 | biostudies-literature | 2018 May

REPOSITORIES: biostudies-literature

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A rapid and nondestructive protocol for whole-mount bone staining of small fish and Xenopus.

Sakata-Haga Hiromi H Uchishiba Maimi M Shimada Hiroki H Tsukada Tsuyoshi T Mitani Mayumi M Arikawa Tomohiro T Shoji Hiroki H Hatta Toshihisa T

Scientific reports 20180510 1

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 c ...[more]

PMID: 29748567

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Dataset Information

A rapid and nondestructive protocol for whole-mount bone staining of small fish and Xenopus.

Publications

A rapid and nondestructive protocol for whole-mount bone staining of small fish and Xenopus.

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