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共聚焦激光扫描显微镜 (CLSM) 作为一种新的工具,用于对新采集和博物馆凭证标本的形态学特征进行研究,这些标本属于 1863 年分类的棘头虫目 (扁形动物:绦虫纲)。

Confocal laser scanning microscopy (CLSM) as a new tool for morphological characterisation of both newly collected and museum voucher specimens of the Trypanorhyncha Diesing, 1863 (Platyhelminthes: Cestoda).

机构信息

Aquaculture and Sea-Ranching, Faculty of Agricultural and Environmental Science, University of Rostock, Rostock, Germany.

Institute of Biosciences, General and Systematic Zoology, University of Rostock, Rostock, Germany.

出版信息

Folia Parasitol (Praha). 2022 Nov 10;69:2022.026. doi: 10.14411/fp.2022.026.

Abstract

Taxonomic issues within Trypanorhyncha, e.g., the inaccurate light microscopic visualisation of the hook patterns, are solvable by confocal laser scanning microscopy (CLSM). We applied CLSM imaging to study Trygonicola macropora (Shipley et Hornell, 1906) and Dollfusiella michiae (Southwell, 1929) from Neotrygon caeruliopunctata Last, White et Séret from Bali, Indonesia. To illustrate the strength and limitations of CLSM, images of Otobothrium cysticum (Mayer, 1842) and Symbothriorhynchus tigaminacantha Palm, 2004, both permanent mounts from a collection, were also processed. The CLSM created image stacks of many layers, and edited with IMARIS Software, these layers resulted in three-dimensional images of the armature patterns and internal organs of both species. BABB (benzylalcohol and benzylbenzuolate) clearing was applied to T. macropora. We conclude that trypanorhynch cestodes stained with Mayer-Schuberg's acetic carmine permanently mounted in Canada balsam are suitable for CLSM, allowing detailed analyses of museum type-material as well as freshly collected and processed worms. BABB resulted in imaging the testes in detail, suggesting other stains to be used for CLSM in trypanorhynch cestode research. Application of CLSM for studies of other cestode groups is highly recommended.

摘要

在 Trypanorhyncha 中,分类学问题,例如钩子模式的不准确的光学显微镜可视化,可通过共聚焦激光扫描显微镜 (CLSM) 解决。我们应用 CLSM 成像技术研究了来自印度尼西亚巴厘岛的 Neotrygon caeruliopunctata Last、White 和 Séret 的 Trygonicola macropora (Shipley et Hornell, 1906) 和 Dollfusiella michiae (Southwell, 1929)。为了说明 CLSM 的优势和局限性,我们还对永久标本 Otobothrium cysticum (Mayer, 1842) 和 Symbothriorhynchus tigaminacantha Palm, 2004 的图像进行了处理。CLSM 生成了许多层的图像堆栈,并使用 IMARIS 软件进行编辑,这些层生成了这两种物种的骨架模式和内部器官的三维图像。我们对 T. macropora 应用了 BABB(苯甲醇和苯甲酸苄酯)清除法。我们得出的结论是,用 Mayer-Schuberg 的醋酸卡红永久固定在加拿大香脂中的棘头虫绦虫可用于 CLSM,允许对博物馆类型材料以及新鲜采集和处理的蠕虫进行详细分析。BABB 成功地详细成像了睾丸,这表明在棘头虫绦虫研究中应使用其他染色剂进行 CLSM。强烈建议将 CLSM 应用于其他绦虫类群的研究。

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