A 5-color flow cytometric method for extended 8-part leukocyte differential.
Microscopic leukocyte differentials display many drawbacks. Several single 5 to 8-color tubes using multiparameter flow cytometry (MFC) are able to provide extended differentials with sequential gating-based analysis strategies. We investigated a new 5-color MFC method to perform an extended 8-part differential with a simplified gating strategy.



Whole blood was stained with a combination of antibodies including HLA-DR-FITC/CD19-PE/CD45-ECD/CD16-PC5 + CD71-PC5/CD5-PC7.



An original approach was developed to exclude debris and straightforwardly gate the cells to identify sixteen populations. Strong correlations were obtained with the analyzer for neutrophils, lymphocytes, monocytes, and eosinophils (R2>0.9). Abnormal cells, such as immature granulocytes and blast cells were identified with a good sensitivity and excellent correlation against cytomorphologic review (R2=0.66 and 0.99, respectively). The choice of HLA-DR and CD5 improved specificity for the identification of activated T-lymphocytes and some lymphoid neoplastic cells, respectively.



Here a new cytometric differential is proposed with a robust gating strategy which may be used even by unskilled cytometrists and can be easily automated. © 2017 International Clinical Cytometry Society.
Service d'Hématologie Biologique, CHU de Dijon, Dijon, France.
Julien
J
Guy
Département d'Hématologie et d'Immunologie Biologiques, Hôpital Henri Mondor, APHP, UPEC, INSERM U955 IMRB, Créteil, France.
Orianne
O
Wagner-Ballon
Service d'Hématologie Biologique, CHU de Dijon, Dijon, France.
Olivier
O
Pages
Service d'Hématologie Biologique, CHU de Dijon, Dijon, France.
François
F
Bailly
Service d'Hématologie Biologique, CHU de Dijon, Dijon, France.
Jessica
J
Borgeot
Service d'Hématologie Biologique, CHU de Nantes, Nantes, France.
Marie-C
MC
Béné
Service d'Hématologie Biologique, CHU de Dijon, Dijon, France.
Marc
M
Maynadié
D000906:Antibodies / Q000737:chemistry* / Q000378:metabolism; D000918:Antibody Specificity; D015703:Antigens, CD / Q000276:immunology / Q000378:metabolism; D018941:Antigens, CD19 / Q000276:immunology / Q000378:metabolism; D015415:Biomarkers / Q000378:metabolism; D005434:Flow Cytometry / Q000379:methods*; D005456:Fluorescent Dyes / Q000737:chemistry*; D058851:GPI-Linked Proteins / Q000276:immunology / Q000378:metabolism; D015870:Gene Expression; D006684:HLA-DR Antigens / Q000276:immunology / Q000378:metabolism; D006801:Humans; D016130:Immunophenotyping / Q000379:methods*; D007938:Leukemia / Q000175:diagnosis / Q000276:immunology / Q000473:pathology; D017493:Leukocyte Common Antigens / Q000276:immunology / Q000378:metabolism; D007958:Leukocyte Count; D007962:Leukocytes / Q000145:classification* / Q000276:immunology / Q000473:pathology; D008223:Lymphoma / Q000175:diagnosis / Q000276:immunology / Q000473:pathology; D009190:Myelodysplastic Syndromes / Q000175:diagnosis / Q000276:immunology / Q000473:pathology; D017452:Receptors, IgG / Q000276:immunology / Q000378:metabolism; D011990:Receptors, Transferrin / Q000276:immunology / Q000378:metabolism; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D000906:Antibodies; D015703:Antigens, CD; D018941:Antigens, CD19; D015415:Biomarkers; C000604811:CD19 molecule, human; C068126:CD71 antigen; C428200:FCGR3B protein, human; D005456:Fluorescent Dyes; D058851:GPI-Linked Proteins; D006684:HLA-DR Antigens; D017452:Receptors, IgG; D011990:Receptors, Transferrin; D017493:Leukocyte Common Antigens; C516769:PTPRC protein, human
flow cytometry; leukocyte differential; reference method