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塑料包埋两栖类胚胎和幼体的组织学

Histology of plastic embedded amphibian embryos and larvae.

作者信息

Kurth Thomas, Weiche Susanne, Vorkel Daniela, Kretschmar Susanne, Menge Anja

机构信息

Center for Regenerative Therapies Dresden, Dresden University of Technology, Tatzberg 47/49, Dresden, Germany.

出版信息

Genesis. 2012 Mar;50(3):235-50. doi: 10.1002/dvg.20821. Epub 2011 Dec 27.

Abstract

Amphibians including the South African clawed frog Xenopus laevis, its close relative Xenopus tropicalis, and the Mexican axolotl (Ambystoma mexicanum) are important vertebrate models for cell biology, development, and regeneration. For the analysis of embryos and larva with altered gene expression in gain-of-function or loss-of-function studies histology is increasingly important. Here, we discuss plastic or resin embedding of embryos as valuable alternatives to conventional paraffin embedding. For example, microwave-assisted tissue processing, combined with embedding in the glycol methacrylate Technovit 7100, is a fast, simple, and reliable method to obtain state-of-the-art histology with high resolution of cellular details in less than a day. Microwave-processed samples embedded in Epon 812 are also useful for transmission electron microscopy. Finally, Technovit-embedded samples are well suited for serial section analysis of embryos labeled either by whole-mount immunofluorescence, or with tracers such as GFP or fluorescent dextrans. Therefore, plastic embedding offers a versatile alternative to paraffin embedding for routine histology and immunocytochemistry of amphibian embryos.

摘要

两栖动物,包括南非爪蟾非洲爪蟾、其近亲热带爪蟾以及墨西哥钝口螈,是细胞生物学、发育和再生研究中重要的脊椎动物模型。在功能获得或功能丧失研究中,对于分析基因表达改变的胚胎和幼体而言,组织学变得越来越重要。在此,我们讨论胚胎的塑料或树脂包埋,这是传统石蜡包埋的宝贵替代方法。例如,微波辅助组织处理,结合在甲基丙烯酸乙二醇酯Technovit 7100中包埋,是一种快速、简单且可靠的方法,能在不到一天的时间内获得具有高分辨率细胞细节的先进组织学结果。包埋在Epon 812中的微波处理样品也可用于透射电子显微镜检查。最后,Technovit包埋的样品非常适合对通过全胚胎免疫荧光或用GFP或荧光葡聚糖等示踪剂标记的胚胎进行连续切片分析。因此,对于两栖动物胚胎的常规组织学和免疫细胞化学研究,塑料包埋提供了一种通用的石蜡包埋替代方法。

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