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使用高内涵成像与分析技术测量细胞健康状况和细胞毒性的工具。

Tools to Measure Cell Health and Cytotoxicity Using High Content Imaging and Analysis.

作者信息

Mandavilli Bhaskar S, Aggeler Robert J, Chambers Kevin M

机构信息

Thermo Fisher Scientific, 29851 Willow Creek Road, Eugene, OR, 97402, USA.

出版信息

Methods Mol Biol. 2018;1683:33-46. doi: 10.1007/978-1-4939-7357-6_3.

Abstract

High content screening (HCS)-based multiparametric measurements are very useful in early toxicity testing and safety assessment during drug development, and useful in evaluating the impact from new food supplements and environmental toxicants. Mitochondrial membrane potential, plasma membrane permeability, oxidative stress, phosphoplipidosis, and steatosis are a few of the important markers routinely studied for the assessment of drug-induced liver injury and toxicity. Mitochondrial dysfunction leads to oxidative stress and cell death. Liver injury from drug-induced phospholipidosis and steatosis is routinely studied in hepatotoxicity investigations to determine the risk factors and fate of drugs or chemical compounds as some drugs can lead to defects in lipid metabolism and accumulation of lipids in lysosomes. In this chapter, we describe fluorescent reagents and the protocols for the measurement of various parameters such as mitochondrial membrane potential, plasma membrane permeability, oxidative stress, phospholipidosis, and steatosis using high content imaging-based methodologies and instrumentation.

摘要

基于高内涵筛选(HCS)的多参数测量在药物研发过程中的早期毒性测试和安全性评估中非常有用,也有助于评估新型食品补充剂和环境毒物的影响。线粒体膜电位、质膜通透性、氧化应激、磷脂沉积症和脂肪变性是常规用于评估药物性肝损伤和毒性的一些重要标志物。线粒体功能障碍会导致氧化应激和细胞死亡。在肝毒性研究中,通常会研究药物诱导的磷脂沉积症和脂肪变性引起的肝损伤,以确定药物或化合物的风险因素和转归,因为某些药物可能导致脂质代谢缺陷和脂质在溶酶体中的积累。在本章中,我们将介绍荧光试剂以及使用基于高内涵成像的方法和仪器测量各种参数(如线粒体膜电位、质膜通透性、氧化应激、磷脂沉积症和脂肪变性)的实验方案。

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