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A novel flow cytometry‐based method of analyzing Heinz bodies
Author(s) -
Palasuwan D.,
Palasuwan A.,
Charoensappakit A.,
Noulsri E.
Publication year - 2017
Publication title -
international journal of laboratory hematology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.705
H-Index - 55
eISSN - 1751-553X
pISSN - 1751-5521
DOI - 10.1111/ijlh.12576
Subject(s) - heinz body , flow cytometry , chemistry , fluorescence , cytometry , population , microbiology and biotechnology , biochemistry , biology , medicine , physics , hemoglobin , optics , environmental health
Summary Introduction Heinz bodies are important to diagnosing and managing patients. However, microscopic examination of Heinz bodies has several disadvantages, demonstrating the need for a better method. We explored the potential use of flow cytometry to examine Heinz bodies. Methods Whole‐blood samples were collected from patients deficient in G6 PD and healthy volunteers. Acetylphenylhydrazine was used to induce formation of Heinz bodies in red blood cells ( RBC s). Then, RBC s positive for Heinz bodies were examined using a FACSC anto II cytometer. Results RBC s treated with acetylphenylhydrazine formed Heinz bodies and emitted a broad spectrum of fluorescence that could be detected by flow cytometry. The maximum emission of fluorescence was observed at 45 min after the incubation with acetylphenylhydrazine. In addition, the fluorescence emitted was stable for at least 72 h. The flow cytometer could detect the RBC s positive for Heinz bodies even if they made up as little as 0.1% of the total RBC population. Furthermore, the percentage and number, respectively, of RBC s positive for Heinz bodies in G6 PD ‐deficient patients and normal donors exhibited a mean ± standard deviation ( SD ) of 68.9 ± 27.5 vs . 50.9 ± 28.6 and 96 014 ±35 732 cells/ μ L vs . 74 688 ± 36 514 cells/ μ L. Conclusion Heinz bodies induced by acetylphenylhydrazine emit fluorescence, and this fluorescence could be examined using flow cytometry. Our study suggests the potential use of the developed method to investigate the formation of Heinz bodies in clinical samples.