A rapid and simple procedure for the whole-mount bone staining of small fish

ABSTRACT

The cartilage and bone structure has provided insightful knowledge about evolution and ecology of fish, which is an important component of biological oceanography. However, the whole-body bone staining is a lengthy and complicated process that typically takes five days to several months, and the improvement of the conventional method has been one of the important issues in this field. Here we report a quick and easy whole-mount bone staining method for small fish, in which a newly designed fixative is applied. Compared to conventional methods, this novel protocol is a straightforward process that could be adopted for small estuarine fish and other small vertebrates.

KEYWORDS:

• Skeleton system
• small estuarine fish
• Thailand
• whole-mount bone staining

Acknowledgements

We thank to acknowledge the Marine Science and Environment, Faculty of Science and Fisheries Technology, Rajamangala University of Technology Srivijaya, Trang campus for laboratory facilities and chemical reagents.

Disclosure statement

No potential conflict of interest was reported by the author(s).

Authors’ contributions

SS was responsible for the overall steps for manuscript study design, data acquisition, and analysis. NY, CS and SS focused on the collection of specimens and experimental verification. CS, SK, KA, JK, PP, AI, PB, KN, AK, GK and SS were responsible for the integrity of the entire study and manuscript review. All authors contributed to the article and approved the submitted version.

Data availability statement

The authors confirm that the data supporting the findings of this study are available within the article and its supplementary materials.

Ethical approval

All procedures in the present study conformed to the guidelines for animal experiments and were approved by the Institutional Animal Care and Use Committee of the Rajamangala University of Technology Srivijaya, Trang Campus (ID#IAC 13-04-64).