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pISSN 2950-9114 eISSN 2950-9122
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Original Article

Lab Med Qual Assur 2024; 46(2): 87-95

Published online June 30, 2024


Copyright © Korean Association of External Quality Assessment Service.

Effect of Whole Blood Fixatives on Cell Fraction and Immunophenotypic Marker Stability in Bone Marrow Specimens Examined by Flow Cytometric Analyses

Woo Yong Shin1 , Hae In Bang2 , Jung-Ah Kim2 , Jieun Kim2 , Rojin Park2 , Jeong Won Shin2 , and Tae Youn Choi2

1Department of Laboratory Medicine, CHA Gangnam Medical Center, CHA University School of Medicine; 2Department of Laboratory Medicine, Soonchunhyang University Seoul Hospital, Seoul, Korea

Correspondence to:Hae In Bang
Department of Laboratory Medicine, Soonchunhyang University Seoul Hospital, 59 Daesagwan-ro, Yongsan-gu, Seoul 04401, Korea
Tel +82-2-709-9430
E-mail genuine43@schmc.ac.kr

Received: November 23, 2023; Revised: March 12, 2024; Accepted: March 13, 2024

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.


Background: Since the clinical application of flow cytometry, many clinical laboratories have utilized this method for diagnosing hematologic malignancies. However, delays in testing can occur due to various reasons. To address this issue, whole blood fixatives are occasionally administered. Hence, this study aimed to determine the impact of applying whole blood fixative on bone marrow specimens.
Methods: Nine samples without lymphoma/leukemia bone marrow involvement were examined. Flow cytometry was performed using 17 common markers. The samples were stored at room temperature (RT) and 4°C without fixative treatment, stored at 4°C after TransFix (Cytomark, UK) treatment, and stored at RT after Cyto-Chex (Streck, USA) treatment. Subsequently, the samples were divided into groups and examined. A total of 13 tests were conducted on each sample for up to 5 days.
Results: The neutrophil and monocyte fractions improved when the samples were stored at 4°C, while no significant difference was observed in the lymphocyte fractions. The fluorescence intensity of the markers varied depending on the marker and conditions, with the most stable markers observed when stored in TransFix at 4°C, followed by storage at 4°C, CytoChex RT, and RT.
Conclusions: The use of fixative on bone marrow specimens maintained the stability of markers during delayed testing. Both fixatives are more effective in preserving marker intensity and cell fractions compared with RT storage. Refrigeration and the use of fixatives may be beneficial for examinations delayed beyond 72 hours.

Keywords: Flow cytometry, Fixatives, Preservative, Bone marrow, Stability

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