Flow Cytometry
lower end’ of the capacity scales in a typical drug screening laboratory where one to two million compounds can be profiled in just a few weeks, but further improvements are in progress (eg, 1536 well sampling). Other important, platform inde- pendent, factors require consideration, such as assay costs and cell supply, particularly the avail- ability of primary human cells.
Despite the significant advances in sample throughput, managing and interpreting complex multi-parametric data remains a challenge. Traditional flow cytometry data analysis tools are not designed to meet this problem, as the typical software package is a complex tool designed for universal use, not to handle high capacity plates, eg, 384 or more wells. Even with 96-well plates, most flow-cytometry analysis approaches are inef- ficient. Better computational analysis tools are required to handle the quantity and complexity of the data. Combining the increased quantity of data from simultaneously measured biomarkers with data collected following cell population perturba- tion after exposure to drugs or other factors (growth factors, etc) may require a multifactorial approach to data analysis, where complex response patterns are generated from heteroge- neous populations. Speciality custom-tailored informatics solutions designed with flow cytome- try in mind will be required, such as the use of Cytobank and advanced visualisation tools such as spanning-tree progression analysis of density-nor- malised events (SPADE)16 or PlateAnalyzer17.
Acknowledgements
The authors would like to acknowledge Poonam Shah and Metul Patel for their scientific contri- butions and Stuart Baddeley (Director, Screening & Compound Profiling, UK) for expert opinion and input.
DDW
Dr Steve Ludbrook is currently a Section Head in Screening & Compound Profiling, Platform Technologies and Science at GlaxoSmithKline in Stevenage, UK. He has worked for more than 15 years in various areas in the target validation through to candidate selection phase, with a cur- rent focus on the use of phenotypic assays in drug discovery.
Dr Rob Jepras is an Investigator in the Screening and Compound Profiling Department at GlaxoSmithKline in Stevenage, UK. He has more than 20 years’ experience at GSK, with a particu- lar focus on high content technologies and primary cell biology in drug discovery.
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