Ann Lab Med 2019; 39(2): 141-149  https://doi.org/10.3343/alm.2019.39.2.141
Development of a Novel Flow Cytometry-Based System for White Blood Cell Differential Counts: 10-color LeukoDiff
Dongjin Park, M.D.1, Jiyoung Chang, M.D.1, Jimin Kahng, M.D.1, Hunhee Park, Ph.D.2, Irene Jo, M.D.1, Yonggoo Kim, M.D.1, and Kyungja Han, M.D.1
1Department of Laboratory Medicine, College of Medicine, The Catholic University of Korea, Seoul, Korea; 2Department of Clinical Laboratory Science, Ansan University, Ansan, Korea
Corresponding author: Kyungja Han, M.D. https://orcid.org/0000-0003-4960-3403
Department of Laboratory Medicine, College of Medicine, Seoul St. Mary’s Hospital, The Catholic University of Korea, 222 Banpo-daero, Seocho-gu, Seoul 07345, Korea
Tel: +82-2-2258-1644
Fax: +82-2-2258-1719
E-mail: hankja@catholic.ac.kr
Received: December 6, 2017; Revised: May 4, 2018; Accepted: August 28, 2018; Published online: March 1, 2019.
© Korean Society for Laboratory Medicine. All rights reserved.

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
Abstract
Background: Flow cytometry (FCM) is commonly used to identify many cell populations. We developed a white blood cell (WBC) differential counting system for detecting abnormal cells using FCM incorporating 10 colors and 11 antibodies in a single tube, called “10-color LeukoDiff,” and evaluated its performance.
Methods: Ninety-one EDTA-anti-coagulated peripheral blood samples from 76 patients were analyzed using 10-color LeukoDiff. We compared 10 color LeukoDiff results with the results of manual differential count (manual diff). WBCs were classified into 17 cell populations: neutrophils, total lymphocytes, T lymphocytes, B lymphocytes, CD5 and CD19 co-expressing lymphocytes, natural killer cells, total monocytes, 16+ monocytes, eosinophils, immature granulocytes, basophils, myeloblasts, B-blasts, T-blasts, myeloid antigen-positive B-blasts, CD19- plasma cells, and 19+ plasma cells.
Results: The correlations between the 10-color LeukoDiff and manual diff results were strong (r>0.9) for mature neutrophils, lymphocytes, eosinophils, immature granulocytes, and blasts and moderate for monocytes and basophils (r=0.86 and 0.74, respectively). There was no discrepancy in blast detection between 10-color LeukoDiff and manual diff results. Furthermore, 10-color LeukoDiff could differentiate the lineage of the blasts and separately count chronic lymphocytic leukemic cells and multiple myeloma cells.
Conclusions: The 10-color LeukoDiff provided an accurate and comprehensive WBC differential count. The most important ability of 10-color LeukoDiff is to detect blasts accurately. This system is clinically useful, especially for patients with hematologic diseases, such as acute leukemia, chronic lymphocytic leukemia, and multiple myeloma. Application of this system will improve the development of FCM gating strategy designs.
Keywords: Flow cytometry, Manual differential count, 10-color LeukoDiff, Blasts, Immature granulocytes



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