High Throughput Screening Readouts
There are many types of high throughput readouts which may be used for screening. Determining which is most appropriate to your screen is usually a balance between data generation and cost per screen.
At the top end of readouts are the high content imaging screens which produce large volumes of content rich information allowing sub-cellular level interrogation of phenotypic change within a cellular population across multiple treatments, time points and/or populations. The data generated is extremely rich in content and allow multiple analyses to be applied once the initial readouts have been compiled.This level of screening can be used to directly assess phenotypic changes such as protein level expression or nuclear translocation within a cell and may be considered to be a more directive form of read out.
Pheontypic assays are rapidly becoming the norm in target discovery, as complex interrogations can be applied to the data sets over time, gradually building up a very complex picture of how the biological active affects a given phenotype.
At the lower end of the scale (in terms of content generated) are simple format biochemical readouts, these usually take the form of a reprted gene assay or biochemmical conversion assays. These are generally considered to be a secondary approximation read. These types of read can be incredibly useful in that they are usually low cost, simple format assays which may be used to determine specific protein acitvation/inactivation.
These types of assay are also used in the TDI as a marker of cellular toxicity during an assay and provide valuable information about cell health and number throughout an assay.
It is important to clearly define the hypothesis being tested to determine the most appropriate read out for your screen.
The TDi is well placed to provide advice on readout selection as we have considerable experience in a wide range of assay platforms.