Proliferation Assays
Titration of Paclitaxel
Quantifying cellular proliferation is one of the most fundamental means for monitoring cells in culture. Proliferation assays are vital in pre-clinical drug discovery for specific therapeutic target areas, such as oncology, and for in-vitro cytotoxicity assays.
Many techniques have been developed to measure cellular proliferation including: quantifying DNA synthesis (3H-Thymidine, *BrdU, DNA dye staining), intracellular reduction-oxidation (1MTT, 2XTT, 3MTS, 4alamarBlue™), and antigen detection (ELISA). All of these techniques can be performed in a microplate format and, as such, are extendable to high-throughput assays. However, these techniques suffer from several disadvantages. The IncuCyte™ provides a novel method for the quantification of cellular proliferation which eliminates many disadvantages associated with currently available assay protocols.
The strength of the IncuCyte™ platform lies in its ability to generate kinetic growth curves directly from phase contrast imagery. Furthermore, the kinetic images are generated inside your incubator, eliminating the need to remove the cells from their controlled environment. This non-invasive, label-free approach offers unique insights into the kinetic dimension of compound effects not revealed by traditional "end-point" assays.
Cellular proliferation depends on many variables including both seeding density and cell type. As such, seeding densities must be adjusted to generate consistent growth curves that, once established, are reproducible. IncuCyte™ can be used to establish and maintain optimized seeding parameters as well as to track consistency of the resulting proliferation curves.
The kinetic assay provided by IncuCyte™ can also help determine compound potency. Fixed "end-point" protocols can generate different results depending on the lag time required before the compound affects proliferation. A full-kinetic profile can help elucidate these discrepancies as well as the mechanism of action of the compound.
Use IncuCyte™ to:
- Elucidate time-dependent action of compounds
- Optimize media, substrate and growth conditions
- Optimize seeding densities
- Determine the optimum sampling window for an end-point HTS assay
