title: A low-volume platform for cell-respirometric screening based on quenched-luminescence oxygen sensing.
abstract: 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. Sealable 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 microl volume has been demonstrated.
authors.affiliation: National Microelectronics Research Centre, Lee Maltings, Prospect Row, Cork, Ireland.
authors.forename: John
authors.initials: J
authors.lastname: Alderman
authors.forename: James
authors.initials: J
authors.lastname: Hynes
authors.forename: Suzanne M
authors.initials: SM
authors.lastname: Floyd
authors.forename: Jan
authors.initials: J
authors.lastname: Krüger
authors.forename: Rosemary
authors.initials: R
authors.lastname: O'Connor
authors.forename: Dmitri B
authors.initials: DB
authors.lastname: Papkovsky
mesh_terms: D015374:Biosensing Techniques / Q000295:instrumentation; D019069:Cell Respiration / Q000502:physiology; D002470:Cell Survival / Q000502:physiology; D049408:Luminescent Agents; D008163:Luminescent Measurements; D010100:Oxygen / Q000032:analysis*; D013997:Time Factors
chemical_list: D049408:Luminescent Agents; D010100:Oxygen