A 5‐color flow cytometric method for extended 8‐part leukocyte differential. Issue 6 (5th April 2017)

• Record Type: Journal Article
• Authors: Guy, Julien; Wagner‐Ballon, Orianne; Pages, Olivier; Bailly, François; Borgeot, Jessica; Béné, Marie‐C; Maynadié, Marc
• Is Part Of: Cytometry. Volume 92:Issue 6(2017)
• Page Start: 498
• Page End: 507
• Publication Date: 2017-04-05
• Subjects: leukocyte differential -- flow cytometry -- reference method; Flow cytometry -- Diagnostic use -- Periodicals; Cytodiagnosis -- Periodicals; 616.07582
• DOI: 10.1002/cyto.b.21524
• Languages: English
• ISSNs: 1552-4949

Abstract

Abstract : Objectives: 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. Methods: Whole blood was stained with a combination of antibodies including HLA‐DR‐FITC/CD19‐PE/CD45‐ECD/CD16‐PC5 + CD71‐PC5/CD5‐PC7. Results: 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 ( R 2 >0.9). Abnormal cells, such as immature granulocytes and blast cells were identified with a good sensitivity and excellent correlation against cytomorphologic review ( R 2 =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. Conclusions: 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