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KMID : 0366220150500020109
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Korean Journal of Hematology
2015 Volume.50 No. 2 p.109 ~ p.112
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Stability of eosin-5’-maleimide dye used in flow cytometric analysis for red cell membrane disorders
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Mehra Simmi
Tyagi Neetu
Dorwal Pranav
Pande Amit
Jain Dharmendra
Sachdev Ritesh
Raina Vimarsh
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Abstract
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Background: The eosin-5’-maleimide (EMA) binding test using flow cytometry is a common method to measure reduced mean channel fluorescence (MCF) of EMA-labeled red blood cells (RBCs) from patients with red cell membrane disorders. The basic principle of the EMA-RBC binding test involves the covalent binding of EMA to lysine-430 on the first extracellular loop of band 3 protein.
Methods: In the present study, the MCF of EMA was analyzed for samples derived from 12 healthy volunteers (controls) to determine the stability (i.e., the percentage decrease in fluorescence) of EMA over a period of 1 year.
Results: Comparison of periodical MCF readings over time, that is, at 2-month intervals, showed that there were no significant changes in mean channel fluorescence for up to 6 months; however, there was a significant decrease in MCF at 8 months.
Conclusion: For optimal dye utilization, EMA remained stable only for up to 6 months. Therefore, we recommend reconstitution of the dye every 6 months when implementing this test and storage at -80oC in dark conditions.
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KEYWORD
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Flow Cytometry, Red Blood cells, Hereditary spherocytosis
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