Fluorescence Intensity Normalisation Correcting for Time Effects in Large-Scale Flow Cytometric Analysis.pdfVIP

Hindawi Publishing Corporation Advances in Bioinformatics Volume 2009, Article ID 476106, 6 pages doi:10.1155/2009/476106 Research Article Fluorescence Intensity Normalisation: Correcting for Time Effects in Large-Scale Flow Cytometric Analysis Calliope A. Dendrou, Erik Fung, Laura Esposito, John A. Todd, Linda S. Wicker, and Vincent Plagnol Juvenile Diabetes Research Foundation/Wellcome Trust Diabetes and Inflammation Laboratory, Department of Medical Genetics, Cambridge Institute for Medical Research, University of Cambridge, Cambridge CB2 0XY, UK Correspondence should be addressed to Vincent Plagnol, vincent.plagnol@cimr.cam.ac.uk Received 28 April 2009; Revised 31 July 2009; Accepted 23 August 2009 Recommended by Ryan R. Brinkman A next step to interpret the findings generated by genome-wide association studies is to associate molecular quantitative traits with disease-associated alleles. To this end, researchers are linking disease risk alleles with gene expression quantitative trait loci (eQTL). However, gene expression at the mRNA level is only an intermediate trait and flow cytometry analysis can provide more downstream and biologically valuable protein level information in multiple cell subsets simultaneously using freshly obtained samples. Because the throughput of flow cytometry is currently limited, experiments may need to span over several weeks or months to obtain a sufficient sample size to demonstrate genetic association. Therefore, normalisation methods are needed to control for technical variability and compare flow cytometry data over an extended period of time. We show how the use of normalising fluorospheres improves the repeatability of a cell surface CD25-APC mean fluorescence intensity phenotype on CD4+ memory T cells. We inve