Increasing output, reducing time, and improving accuracy of laboratory assays can be achieved by various new technologies that encompass multiplexing, low volumes, high-throughput screening, and advanced software for collection and analysis of large datasets.
Multiplexing provides the ability to obtain multiple datasets from a single assay condition.
The key benefit of multiplexing is that you gain a better and more accurate understanding of a complex system by measuring the sum of the multiple parts. If only a single activity is measured, the effect of experimental treatment on cell outcome could be overlooked. Multiplexing can also include the measurement of multiple cellular properties from a single marker, for example some nucleic acid dyes can provide information about more than one aspect of cell health. Ratios of parameters can also be calculated enabling normalization of data, for example, the ratio of a cell’s activity to the cell number in that well.
In setting up multiplexing assays, accurate optimization is required to reduce overlap between multiple signals and provide compatibility under similar conditions. There is also the requirement for equipment that can detect and analyze multiple signals in a time-dependent way. As the development of these technologies advances and more detection products become available, increasingly less time is required to optimize assays, creating a system that is more accessible and suitable for a larger number of applications.
Various detection systems are available that work on a planar or suspension system depending on the specific target required. Established chemiluminescent detection technologies are readily being replaced by fluorescent-based systems due to the advancement of a greater number of fluorophores that can be used simultaneously.
In combination with more sophisticated plate readers capable of imaging whole cells in high-throughput mode, many industries have been revolutionized, particularly drug discovery.
In this article, we discuss the identification of several hepatotoxicity effects of three commercially available compounds on a liver cell line using an advanced multiplex assay.
Unpredicted side-effects and toxicity issues limit or prevent a candidate molecule being taken to market. This can often occur very late in the testing phases, thereby incurring substantial time and cost factors for the companies involved.