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Alderman, J.,Hynes, J.,Floyd, S. M.,Kruger, J.,O'Connor, R.,Papkovsky, D. B.;
A low-volume platform for cell-respirometric screening based on quenched-luminescence oxygen sensing
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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.
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