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Kaido M, Takagi Y, Kono M, et al. Investigation of morphological changes for the discrimination of nucleated red blood cells and other leukocytes in Sysmex XN hematology analyzer scattergrams using transmission electron microscopy. Pract Lab Med. 2017;8:70-76doi: 10.1016/j.plabm.2017.05.001.
Kaido, M., Takagi, Y., Kono, M., Nakazawa, F., Yamamoto, S., Wada, A., & Morikawa, T. (2017). Investigation of morphological changes for the discrimination of nucleated red blood cells and other leukocytes in Sysmex XN hematology analyzer scattergrams using transmission electron microscopy. Practical laboratory medicine, 870-76. https://doi.org/10.1016/j.plabm.2017.05.001
Kaido, Masako, et al. "Investigation of morphological changes for the discrimination of nucleated red blood cells and other leukocytes in Sysmex XN hematology analyzer scattergrams using transmission electron microscopy." Practical laboratory medicine vol. 8 (2017): 70-76. doi: https://doi.org/10.1016/j.plabm.2017.05.001
Kaido M, Takagi Y, Kono M, Nakazawa F, Yamamoto S, Wada A, Morikawa T. Investigation of morphological changes for the discrimination of nucleated red blood cells and other leukocytes in Sysmex XN hematology analyzer scattergrams using transmission electron microscopy. Pract Lab Med. 2017 May 10;8:70-76. doi: 10.1016/j.plabm.2017.05.001. eCollection 2017 Aug. PMID: 28856231; PMCID: PMC5575374.
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Pract Lab Med. 2017 May 10;8:70-76. doi: 10.1016/j.plabm.2017.05.001. eCollection 2017 Aug.

Investigation of morphological changes for the discrimination of nucleated red blood cells and other leukocytes in Sysmex XN hematology analyzer scattergrams using transmission electron microscopy.

Practical laboratory medicine

Masako Kaido, Yuri Takagi, Mari Kono, Fumie Nakazawa, Shiori Yamamoto, Atsushi Wada, Takashi Morikawa

Affiliations

  1. Scientific Research, Scientific Affairs, Sysmex Corporation, 1-3-2 Murotani, Nishi-ku, Kobe 651-2241, Japan.

PMID: 28856231 PMCID: PMC5575374 DOI: 10.1016/j.plabm.2017.05.001

Abstract

BACKGROUND: The WNR channel of the XN-Series automated hematology analyzer (Sysmex) counts white blood cells (WBCs) and simultaneously performs a differential counting of basophils and nucleated red blood cells (NRBCs). The detection process involves exposing the cells to WNR-specific reagents containing an acidic detergent and a fluorescent dye and measuring the intensity of the forward scattered light (FSC) and side fluorescence light (SFL).

METHOD: We treated isolated peripheral WBCs and NRBCs with specific reagents and assessed the morphological changes in NRBCs and each leukocyte type using transmission electron microscopy (TEM).

RESULTS: The results from a flow cytometer (FCM) showed that, after exposure to the reagents, basophils appeared on the highest FSC and SFL areas compared to other leukocytes on the WNR scattergram. Owing to the hemolysis of reticulocytes and erythrocytes, NRBCs that survived the reagent treatment could be distinguished by their lower intensity than those of the other leukocytes on the WNR scattergram. We investigated the significance of the relationship between the TEM and FCM results after the reagent treatment.

CONCLUSION: We confirmed that the WNR channel differentiates the blood cells on the WNR scattergram based on differences in the amount of residual cytoplasm and nucleic acids.

Keywords: Automated hematology analyzer; EDTA, ethylenediaminetetraacetic acid; FCM, flow cytometer; FITC, fluorescein isothiocyanate; FSC, forward scattered light; Flow cytometry; Fluorescent intensity; Leukocytes; MACS, magnetic cell sorting; MAS, Matsunami Adhesive Silane; NRBCs; NRBCs, nucleated red blood cells; PE, phycoerythrin; SFL, side fluorescence light; Scatter light intensity; TEM, transmission electron microscopy; Transmission electron microscopy; WBCs, white blood cells

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