A low-volume platform for cell-respirometric screening based on quenched-luminescence oxygen sensing
A low-volume platform for cell-respirometric screening based on quenched-luminescence oxygen sensing
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DOI：
10.1016/j.bios.2003.12.008
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发表时间：
2004-06-15
影响因子：
12.6
通讯作者：
Papkovsky, DB
中科院分区：
文献类型：
--
作者：
Alderman, J;Hynes, J;Papkovsky, DB
细胞活性检测是现代细胞生物学、药物开发和生物技术中的一项重要技术，目前对简单、灵敏和经济有效的筛选方法有很高的需求。我们已经开发了一种基于细胞的筛选分析的新方法和相关工具，该方法基于通过发光猝灭测量细胞中的氧摄取率。与标准96孔板的占地面积相匹配的可扩展微室装置被开发出来，并与长衰变磷光氧探测器一起使用。这些设备允许细胞非侵入性地实时监测细胞呼吸，并对多个样本的细胞活力、药物/效应器作用进行快速、一步的动力学评估。这些分析可以在传统的荧光平板阅读器上进行，它们适用于不同类型的细胞，包括贴壁细胞和慢反应细胞，需要较小的样本量和细胞数量，并适合高通量筛选。已经证明，可以在3微升的体积中监测到300个哺乳动物细胞。(C)2004爱思唯尔B.V.保留所有权利。
Cell viability assays represent an important technology in modern cell biology, drug discovery and biotechnology, where currently there is a high demand for simple, sensitive and cost-effective screening methods. We have developed a new methodology and associated tools for cell-based screening assays, which are based on the measurement of the rates of oxygen uptake in cells by luminescence quenching. Scalable microchamber devices matching the footprint of a standard 96-well plate were developed and used in conjunction with long-decay phosphorescent oxygen probes. These devices permit cell non-invasive, real-time monitoring of cellular respiration and a rapid, one-step, kinetic assessment of multiple samples for cell viability, drug/effector action. These assays can be carried out on conventional fluorescence plate readers, they are suitable for different types of cells, including adherent and slow-respiring cells, require small sample volumes and cell numbers, and are amenable for high throughput screening. Monitoring of as little as 300 mammalian cells in 3 mul volume has been demonstrated. (C) 2004 Elsevier B.V. All rights reserved.