SA Lugovskaya1, FA Dyukov1, EV Naumova1, IYu Bugrov1, AI Kostin2
1 Russian Medical Academy of Postgraduate Education, 2/1 Barrikadnaya str., Moscow, Russian Federation, 125993
2 NV Sklifosovskii Research Institute of Emergency Medicine, 3 Bol’shaya Sukharevskaya sq. Moscow, Russian Federation, 129090
For correspondence: Prof. Svetlana Alekseevna Lugovskaya, MD, PhD, 2/1 Barrikadnaya str., Moscow, Russian Federation, 125993; Tel.: +7(495)945-82-22; e-mail: slugovskaya@mail.ru
For citation: Lugovskaya SA, Dyukov FA, Naumova EV, Bugrov IYu, et al. The Use of Multicolor Flow Cytofluorometry in White Blood Cell Differential: HematoFlow Conception. Clinical oncohematology. 2018;11(4):319–25.
DOI: 10.21320/2500-2139-2018-11-4-319-325
ABSTRACT
Aim. To develop reference intervals for white blood cell subpopulations in peripheral blood using Cytomics FC500 flow cytometer and CytoDiffTM reagent (Beckman Coulter).
Materials & Methods. The trial included the analysis of blood samples of healthy donors (n = 315) using Cytomics FC500 flow cytometer and CytoDiffTM reagent cocktail composed of 6 antibodies in 5 colors (CD36-FITC, CD2-PE, CD294-PE, CD19-ECD, CD16-Cy5, CD45-Cy7) and enabling to count 17 cell populations.
Results. The data obtained by means of multicolor flow cytofluorometry included the reference values of white blood cell populations in peripheral blood. In 1 first-time donor a lymphoproliferative disease was detected. It was subsequently confirmed by immunophenotyping; B-cell chronic lymphocytic leukemia was diagnosed.
Conclusion. Multicolor flow cytofluorometry using CytoDiffTM is considered to be a new step toward an improved WBC differential evaluation aimed mainly at reducing the volume of blood smear analysis using light microscopy at large laboratories, enhancing objectivity, precision and reproducibility of results. WBC differential extended with the count of lymphocyte and monocyte subpopulations can be regarded as modern donor blood screening to detect changes in the pattern of lymphocyte subpopulations as grounds for further examination of donors.
Keywords: multicolor flow cytofluorometry, reference intervals, WBC differential.
Received: March 11, 2018
Accepted: July 5, 2018
REFERENCES
-
Koepke JA, van Assendelft OW, Brindza LJ, et al. Reference leukocyte (WBC) differential count (proportional) and evaluation of instrumental methods. Wayne, PA: CLSI; 2007. pp. 1–35.
-
Rumke CL. Laboratory aids. Variability of results in differential cell counts on blood smears. Triangle. 1960;4:154–8.
-
Novis DA, Walsh M, Wilkinson D, et al. Laboratory Productivity and the Rate of Manual Peripheral Blood Smear Review: A College of American Pathologists Q-Probes Study of 95,141 Complete Blood Count Determinations Performed in 263 Institutions. Arch Pathol Lab Med. 2006;130(5):596–601.
-
Roussel M, Benard C, Ly-Sunnaram B, Fest T. Refining the white blood cell differential: The first flow cytometry routine application. Cytometry. 2010;77A(6):552–63. doi: 10.1002/cyto.a.20893.
-
Roussel M, Davis BH, Fest T, Wood BL. Toward a reference method for leukocyte differential counts in blood: Comparison of three flow cytometric candidate methods. Cytometry. 2012;81A(11):973–82. doi: 10.1002/cyto.a.22092.
-
Elghetany MT, Lacombe F. Physiologic variations in granulocytic surface antigen-expression: Impact of age, gender, pregnancy, race, and stress. J Leuk Biol. 2004;75(2):157–62. doi: 10.1189/jlb.0503245.
-
Fromont P, Bettaieb A, Skouri H, et al. Frequency of the polymorphonuclear neutrophil Fc gamma receptor III deficiency in the French population and its involvement in the development of neonatal alloimmune neutropenia. Blood. 1992;79(8):2131–4.
-
Meddws-Taylor S, Martin DJ, Tiemessen CT. Altered Expression of FcγRIII (CD16) on Polymorphonuclear Neutrophils from Individuals with Human Immunodeficiency Virus Type 1 Disease and Pulmonary Tuberculosis. Clin Diagn Lab Immunol. 1997;4(6):789–91.
-
Faucher J-L, Lacronique-Gazaille C, Frebet E, et al. “6 Markers/5 Colors” Extended White Blood Cell Differential by Flow Cytometry. Cytometry Part A. 2007;71А(11):934–44. doi: 10.1002/cyto.a.20457.
-
Huang CM, Yu LH, Pu CW, et al. Assessment of a five-color flow cytometric assay for verifying automated white blood cell differentials. Chin Med J. 2013;126(4):716–21.
-
Qu C, Wang J, Pu C, et al. Efficiency of Leukocyte Differential Using Flow Cytometry with CytoDiff in Different Workflows. Clin Lab. 2017;63(4):659–68. doi: 10.7754/Clin.Lab.2017.161221.