Abstract for Multiplexing Cell-Based Assays
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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