Dynamic Expression Profiles from Static Cytometry Data Component Fitting and Conversion to Relative, “Same Scale” Values 英文参考文献.docVIP

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Dynamic Expression Profiles from Static Cytometry Data Component Fitting and Conversion to Relative, “Same Scale” Values 英文参考文献.doc

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Dynamic Expression Profiles from Static Cytometry Data Component Fitting and Conversion to Relative, “Same Scale” Values 英文参考文献

DynamicExpressionProfilesfromStaticCytometryData: ComponentFittingandConversiontoRelative,‘‘Same Scale’’Values JayantAvva1,MichaelC.Weis1,R.MichaelSramkoski2,SreeN.Sreenath1,JamesW.Jacobberger2* 1Department of Electrical Engineering and Computer Sciences, Case Western Reserve University, Cleveland, Ohio, United States of America, 2Case Comprehensive CancerCenter,CaseWesternReserveUniversity,Cleveland,Ohio,UnitedStatesofAmerica Abstract Background: Cytometry of asynchronous proliferating cell populations produces data with an extractable time-based featureembeddedinthefrequencyofclustered,correlatedevents.Here,wepresentaspecificcaseofgeneralmethodology forcalculatingdynamicexpressionprofilesofepitopesthatoscillateduringthecellcycleandconversionofthesevaluesto thesamescale. Methods:SamplesofK562cellsfromonepopulationwerelabeledbydirectandindirectantibodymethodsforcyclinsA2 andB1andphospho-S10-histoneH3.Thesameindirectantibodywasusedforbothcyclins.Directlystainedsampleswere counter-stained with 496-diamidino-2-phenylindole and indirectly stained samples with propidium to label DNA. The S phasecyclin expressions fromindirectassays wereused toscaletheexpression ofthecyclins of themulti-variatedirect assay.Booleangatingandtwodimensional,sequentialregionssetonbivariatedisplaysofthedirectlyconjugatedsample datawereusedtountangleandisolateunique,unambiguousexpressionvaluesofthecyclinsalongthefour-dimensional data path through the cell cycle. The median values of cyclins A2 and B1 from each region were correlated with the frequencyofeventswithineachregion. Results: The sequential runs of data were plotted as continuous multi-line linear equations of the form y = [(yi+12yi)/ (xi+12xi)]x+yi2[(yi+12yi)/(xi+12xi)]xi(linebetweenpoints(xi,yi)and(xi+1,yi+1))tocapturethedynamicexpressionprofileof thetwocyclins. Conclusions: Thisspecificapproach demonstrates thegeneralmethodology andprovides aruleset fromwhich thecell cycleexpressionofanyotherepitopescouldbemeasuredandcalculate