%0 Journal Article %1 McLaughlin2008a %A McLaughlin, Bridget E. %A Baumgarth, Nicole %A Bigos, Martin %A Roederer, Mario %A De Rosa, Stephen C. %A Altman, John D. %A Nixon, Douglas F. %A Ottinger, Janet %A Li, Judy %A Beckett, Laurel %A Shacklett, Barbara L. %A Evans, Thomas G. %A Asmuth, David M. %D 2008 %J Cytometry A %K facs tlymphocytes bcells highthroughput %N 5 %P 411--420 %R 10.1002/cyto.a.20556 %T Nine-color flow cytometry for accurate measurement of T cell subsets and cytokine responses. Part II: Panel performance across different instrument platforms. %U http://dx.doi.org/10.1002/cyto.a.20556 %V 73 %X Cellular immune responses elicited by vaccination are complex and require polychromatic analysis to accurately characterize the phenotype and function of rare, responding cells. Technical challenges and a lack of instrument standardization between research sites have limited the application of polychromatic cytometry in multicenter clinical trials. Two previously developed six-color T cell subset immunophenotyping reagent panels deliberately designed to accommodate three additional low frequency functional measurements were compared for their reproducibility of staining across three different flow cytometers. We repeatedly measured similar T cell subset frequencies between the two reagent panels and across the three different cytometers. Spectral overlap reduced sensitivity in two of the three open measurement channels (PE IL-2 and APC IFN gamma) for one reagent combination, particularly in subsets with low cytokine expression. There was no significant interassay variation for measurements across instrument platforms. Careful panel design will identify reagent combinations that minimize spectral spillover into channels reserved for cytokine measurement and comparable results can be achieved using different cytometers, however, it is important to establish standardized quality control procedures for each instrument to minimize variation between cytometers.

@article{McLaughlin2008a, abstract = {Cellular immune responses elicited by vaccination are complex and require polychromatic analysis to accurately characterize the phenotype and function of rare, responding cells. Technical challenges and a lack of instrument standardization between research sites have limited the application of polychromatic cytometry in multicenter clinical trials. Two previously developed six-color T cell subset immunophenotyping reagent panels deliberately designed to accommodate three additional low frequency functional measurements were compared for their reproducibility of staining across three different flow cytometers. We repeatedly measured similar T cell subset frequencies between the two reagent panels and across the three different cytometers. Spectral overlap reduced sensitivity in two of the three open measurement channels (PE [IL-2] and APC [IFN gamma]) for one reagent combination, particularly in subsets with low cytokine expression. There was no significant interassay variation for measurements across instrument platforms. Careful panel design will identify reagent combinations that minimize spectral spillover into channels reserved for cytokine measurement and comparable results can be achieved using different cytometers, however, it is important to establish standardized quality control procedures for each instrument to minimize variation between cytometers.}, added-at = {2012-11-19T22:30:54.000+0100}, author = {McLaughlin, Bridget E. and Baumgarth, Nicole and Bigos, Martin and Roederer, Mario and {De Rosa}, Stephen C. and Altman, John D. and Nixon, Douglas F. and Ottinger, Janet and Li, Judy and Beckett, Laurel and Shacklett, Barbara L. and Evans, Thomas G. and Asmuth, David M.}, biburl = {https://www.bibsonomy.org/bibtex/2bcac73840558c0869ba165e432a34d7d/aorchid}, description = {continues from McLaughlin2008 with comparison of performance}, doi = {10.1002/cyto.a.20556}, file = {:allorejection/CytoA.73.411.pdf:PDF}, groups = {public}, institution = {University of California Davis, Internal Medicine, Infectious Diseases, Davis, CA, USA.}, interhash = {3eff4bc5bec50304b7d45b433f517c03}, intrahash = {bcac73840558c0869ba165e432a34d7d}, journal = {Cytometry A}, keywords = {facs tlymphocytes bcells highthroughput}, language = {eng}, medline-pst = {ppublish}, month = May, number = 5, pages = {411--420}, pmid = {18383309}, timestamp = {2012-11-19T22:30:54.000+0100}, title = {Nine-color flow cytometry for accurate measurement of T cell subsets and cytokine responses. Part II: Panel performance across different instrument platforms.}, url = {http://dx.doi.org/10.1002/cyto.a.20556}, username = {aorchid}, volume = 73, year = 2008 }