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利用尼罗红和油红O染色对线虫脂质丰度进行定量及评估脂质分布

Quantification of Lipid Abundance and Evaluation of Lipid Distribution in Caenorhabditis elegans by Nile Red and Oil Red O Staining.

作者信息

Escorcia Wilber, Ruter Dana L, Nhan James, Curran Sean P

机构信息

Leonard Davis School of Gerontology, University of Southern California.

Leonard Davis School of Gerontology, University of Southern California; Molecular & Computational Biology Section, University of Southern California; Integrative Program for Biological & Genome Sciences, University of North Carolina at Chapel Hill.

出版信息

J Vis Exp. 2018 Mar 5(133):57352. doi: 10.3791/57352.

Abstract

Caenorhabditis elegans is an exceptional model organism in which to study lipid metabolism and energy homeostasis. Many of its lipid genes are conserved in humans and are associated with metabolic syndrome or other diseases. Examination of lipid accumulation in this organism can be carried out by fixative dyes or label-free methods. Fixative stains like Nile red and oil red O are inexpensive, reliable ways to quantitatively measure lipid levels and to qualitatively observe lipid distribution across tissues, respectively. Moreover, these stains allow for high-throughput screening of various lipid metabolism genes and pathways. Additionally, their hydrophobic nature facilitates lipid solubility, reduces interaction with surrounding tissues, and prevents dissociation into the solvent. Though these methods are effective at examining general lipid content, they do not provide detailed information about the chemical composition and diversity of lipid deposits. For these purposes, label-free methods such as GC-MS and CARS microscopy are better suited, their costs notwithstanding.

摘要

秀丽隐杆线虫是研究脂质代谢和能量稳态的一种特殊模式生物。它的许多脂质基因在人类中是保守的,并且与代谢综合征或其他疾病相关。可以通过固定染料或无标记方法来检测该生物体中的脂质积累。像尼罗红和油红O这样的固定染色剂分别是定量测量脂质水平和定性观察脂质在组织中分布的廉价、可靠方法。此外,这些染色剂允许对各种脂质代谢基因和途径进行高通量筛选。此外,它们的疏水性质有利于脂质溶解,减少与周围组织的相互作用,并防止解离到溶剂中。尽管这些方法在检测一般脂质含量方面有效,但它们不能提供有关脂质沉积物的化学成分和多样性的详细信息。出于这些目的,尽管成本较高,但无标记方法如气相色谱-质谱联用(GC-MS)和相干反斯托克斯拉曼散射(CARS)显微镜更适合。

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