Long-term Live Imaging Device for Improved Experimental Manipulation of Zebrafish Larvae

Kayla Huemer; Jayne M. Squirrell; Robert Swader; Kirsten Pelkey; Danny C. LeBert; Anna Huttenlocher; Kevin W. Eliceiri

doi:10.3791/56340

JoVE Journal
Bioengineering

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JoVE Journal Bioengineering

Long-term Live Imaging Device for Improved Experimental Manipulation of Zebrafish Larvae

改善斑马鱼幼虫实验操作的长期活体成像装置

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14:05 min

October 27, 2017

DOI: 10.3791/56340-v

Kayla Huemer*^1,2, Jayne M. Squirrell*³, Robert Swader², Kirsten Pelkey², Danny C. LeBert⁴, Anna Huttenlocher^5,6, Kevin W. Eliceiri^1,2,3

¹Department of Biomedical Engineering,University of Wisconsin-Madison, ²Morgridge Institute for Research,University of Wisconsin-Madison, ³Laboratory for Optical and Computational Instrumentation,University of Wisconsin-Madison, ⁴Cellular and Molecular Pathology Graduate Program,University of Wisconsin-Madison, ⁵Department of Medical Microbiology and Immunology,University of Wisconsin-Madison, ⁶Department of Pediatrics,University of Wisconsin-Madison

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这份手稿描述了 zWEDGI (斑马鱼伤人和诱捕装置的增长和成像), 这是一个条块分割的装置, 旨在定位和抑制斑马鱼幼虫。该设计允许尾部横断面和长期收集高分辨率的荧光显微图像的伤口愈合和再生。

这种成像设备制造的总体目标是创建一个可修改的平台，用于对活斑马鱼幼虫的伤口愈合进行显微分析。这种方法可以帮助回答伤口愈合领域的关键问题，例如胶原纤维重组和细胞反应对受伤后后续组织再生的贡献是什么。这种技术的主要优点是伤口和成像发生在单个设备中，而尾部再生不受设备阻碍。

此外，zWEDGI 的多个功能区室可以独立修改，以适应不同的幼虫阶段和一系列实验方案。首先，根据文本协议使用所需的几何结构和属性对器件的 PDMS 组件进行建模。使用光敏树脂 3D 打印机打印模具后，使用细刷、喷雾瓶中的变性酒精和压缩空气清洁模具。

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生物工程问题 128 伤口愈合斑马鱼多显微镜克制装置幼虫长期成像