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通过光学显微镜和电子显微镜观察秀丽隐杆线虫中的脂滴。

Visualization of lipid droplets in C. elegans by light and electron microscopy.

作者信息

Mak Ho Yi

机构信息

Stowers Institute for Medical Research, Kansas City, Missouri, USA; Department of Molecular and Integrative Physiology, University of Kansas Medical Center, Kansas City, Kansas, USA.

出版信息

Methods Cell Biol. 2013;116:39-51. doi: 10.1016/B978-0-12-408051-5.00003-6.

Abstract

The powerful forward and reverse genetic tools, and emerging sets of biochemical assays for fat metabolites, make Caenorhabditis elegans an attractive model organism for elucidating conserved mechanisms in fat storage. The ability to observe lipid droplets in live animals at single cell resolution offers a unique advantage for studying cellular fat storage in vivo. In this chapter, we describe transgenic technologies for expressing fluorescent lipid droplet marker proteins at near-physiological levels. Methods to visualize these markers using sensitive confocal microscopy systems are detailed. Additional methods for visualizing lipid droplets by transmission electron microscopy and detection of lipid droplet associated proteins by immunoelectron microscopy are described.

摘要

强大的正向和反向遗传工具,以及针对脂肪代谢产物不断涌现的一系列生化检测方法,使得秀丽隐杆线虫成为阐明脂肪储存保守机制的极具吸引力的模式生物。在活的动物中以单细胞分辨率观察脂滴的能力为体内研究细胞脂肪储存提供了独特优势。在本章中,我们描述了用于在接近生理水平表达荧光脂滴标记蛋白的转基因技术。详细介绍了使用灵敏的共聚焦显微镜系统观察这些标记物的方法。还描述了通过透射电子显微镜观察脂滴以及通过免疫电子显微镜检测脂滴相关蛋白的其他方法。

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