A low-volume platform for cell-respirometric screening based on quenched-luminescence oxygen sensing.
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.
Alderman
John
J
National Microelectronics Research Centre, Lee Maltings, Prospect Row, Cork, Ireland.
Hynes
James
J
Floyd
Suzanne M
SM
Krüger
Jan
J
O'Connor
Rosemary
R
Papkovsky
Dmitri B
DB
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
D049408:Luminescent Agents; D010100:Oxygen