Application of an optical clearing reagent "Scale" to fish pathology

2015 Fiscal Year Final Research Report

• Project/Area Number: 25660154
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Aquatic bioproduction science
• Research Institution: Tokyo University of Marine Science and Technology
• Principal Investigator: ENDO Makoto 東京海洋大学, その他部局等, 教授 (80128355)
• Co-Investigator(Kenkyū-buntansha): MAITA Masashi 東京海洋大学, 学術研究院, 教授 (60238839) ; KATAGIRI Takayuki 東京海洋大学, 学術研究院, 助教 (50361811) ; FUTAMI Kunihiko 東京海洋大学, 学術研究院, 助教 (00513459)
• Project Period (FY): 2013-04-01 – 2016-03-31
• Keywords: 組織透明化試薬 / 蛍光標識 / 魚類病原細菌 / 病理組織学 / 分子生物学

Outline of Final Research Achievements

Fish diseases are generally diagnosed by histological and/or molecular biological analysis following the observation by eyes. However, it is difficult to examine the whole body and clarify the localization of pathogens. Here, we investigated whether the recently developed transparent techniques such as Scale, SeeDB, ClearT and CUBIC are applicable to diagnosis of fish diseases.
CUBIC, which is modified from Scale was most effectively transparentized the fish body, and applicable to imaging of GFP and immunostaining. Furthermore, Aeromonas hydrophila labeled with DAPI could be visualized under a fluorescent microscope. Although DNAs were preserved in all transparent specimens and applicable to PCR, RNA was degraded by CUBIC due to containing basic aminoalcohols. On the other hand, RNA was highly preserved in organs treated with SeeDB or ClearT. Our results indicate that proper use of transparent techniques enable whole body diagnosis of the infectious disease in fish.

Free Research Field

環境毒性学