Estimation of Immune Cell Densities in Immune Cell Conglomerates An Approach for High-Throughput Quantification 英文参考文献.docVIP

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Estimation of Immune Cell Densities in Immune Cell Conglomerates An Approach for High-Throughput Quantification 英文参考文献.doc

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Estimation of Immune Cell Densities in Immune Cell Conglomerates An Approach for High-Throughput Quantification 英文参考文献

EstimationofImmuneCellDensitiesinImmuneCell Conglomerates:AnApproachforHigh-Throughput Quantification NielsHalama1,2,InkaZoernig1,AnnaSpille1,KathiWestphal2,PeterSchirmacher3,DirkJaeger1. ,Niels Grabe2. * 1Medical Oncology, National Center for Tumor Diseases, University of Heidelberg, Heidelberg, Germany, 2Hamamatsu Tissue Imaging and Analysis (TIGA) Center, InstituteforMedicalBiometryandInformatics,UniversityofHeidelberg,Heidelberg,Germany,3InstituteofPathology,UniversityofHeidelberg,Heidelberg,Germany Abstract Background:Determiningthecorrectnumberofpositiveimmunecellsinimmunohistologicalsectionsofcolorectalcancer andothertumorentitiesisemergingasanimportantclinicalpredictorandtherapyselectorforanindividualpatient.This taskisusuallyobstructedbycellconglomeratesofvarioussizes.Wehereshowthatatleastincolorectalcancertheinclusion ofimmunecellconglomeratesisindispensableforestimatingreliablepatientcellcounts.Integratingvirtualmicroscopyand imageprocessingprincipallyallowsthehigh-throughputevaluationofcompletetissueslides. Methodology/Principal findings: For such large-scale systems we demonstrate a robust quantitative image processing algorithm for the reproducible quantification of cell conglomerates on CD3 positive T cells in colorectal cancer. While isolatedcells(28to80mm2)arecounteddirectly,thenumberofcellscontainedinaconglomerateisestimatedbydividing theareaoftheconglomerateinthintissuessections(#6mm)bythemedianareacoveredbyanisolatedTcellwhichwe determinedas58mm .WeappliedouralgorithmtolargenumbersofCD3positiveTcellconglomeratesandcomparedthe resultstocellcountsobtainedmanuallybytwoindependentobservers.Whileespeciallyforhighcellcounts,themanual countingshowedadeviationofupto400cells/mm2(41%variation),algorithm-determinedTcellnumbersgenerallylayin betweenthemanuallyobservedcellnumbersbutwithperfectreproducibility. 2 Conclusion: Insummary, werecommendour approach as anobjectiveand robuststrategy for quantifying immune cell densities in immunohistological sectio