Promega Corporation

Multiplexing Cell-Based Assays Scientific Poster

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Abstract

Terry L. Riss, Erika Hawkins, Jim Cali, Rich A. Moravec and Andrew Niles
Promega Corporation, 2800 Woods Hollow Road, Madison WI 53711

Multiplex recording of more than one indicator from the same sample increases the efficiency of assay development and screening. Recent advances in cell-based assay chemistries have made it possible to record multiplex data in a high throughput format using standard plate readers without the need to perform microscopic imaging of individual cells. Identification of novel markers of cell viability and development of detection chemistries that do not kill cells have expanded the possible combinations for multiplexing cell-based assays in microwell plates.

We have developed methods to combine various luminescent and fluorescent cell-based assays in a homogeneous format. We have recorded luciferase reporter gene assays in real time in living cells followed by measuring cell viability or caspase activity. Measuring the number of viable cells at the end of a treatment period is useful to distinguish between a specific down regulation of a reporter gene or nonspecific cytotoxicity. Multiplexed cell viability data also can serve as an internal control to correct for variability in seeding density and differential growth of cells resulting in improvements in the reliability of data. We have also developed methods for simultaneous detection of viable and dead cells in the same sample and have demonstrated multiplexing of homogeneous fluorescent cell viability and luminescent caspase assays in the same well. We will present the results of ongoing efforts to investigate the compatibility of different assay chemistries and detection methods for developing multiplexed homogeneous cell-based assays with sensitivity sufficient for microwell assays.

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  • Part# PS047
  • Printed in USA.

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