Effect of red blood cell lysing agent in leukemia/lymphoma immunophenotyping by flow cytometry

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dc.contributor.authorRahman, Ken_US
dc.contributor.authorKhan, MAen_US
dc.contributor.authorSarkar, MKen_US
dc.contributor.authorBiswas, Sen_US
dc.contributor.authorKumar, Aen_US
dc.contributor.authorSingh, MKen_US
dc.contributor.authorChandra, Den_US
dc.contributor.authorGupta, R.en_US
dc.date.accessioned2025-05-09T06:17:26Z
dc.date.available2025-05-09T06:17:26Z
dc.date.issued2024-03
dc.description.abstractBackground: Getting rid of the red blood cells (RBC) is an integral part of flow cytometry (FCM)-based leukemia/lymphoma immunophenotyping. It is routinely done using an RBC lysing agent. RBC lysing has numerous consequences for the physical and immunophenotypic characteristics of the cell. We aimed to analyse the effect of different RBC lysing agents (with or without paraformaldehyde) on the antibody staining pattern.Methods: Three different RBC lysis preparations were used, namely BD FACSLyse (FL) and BD PharmLyse (PL), and a laboratory-prepared (home-brewed, HB) ammonium chloride-based reagent. A lyse-stain-wash protocol was used to prepare the cells, which were stained usinga single tube of five-color antibodies (CD20-BV605, CD8-BV510, CD3-FITC, CD4-CD19-PeCy7, and CD45-APCR700). The effects of different lysings on different cellular characteristics, including the median fluorescence intensity (MFI) of CD45 in lymphocytes, CD3 in T lymphocytes, CD19 in B lymphocytes, CD20 in B lymphocytes, CD4 in T helper cells, CD8 in cytotoxic T cells, and CD4 in monocytes, were noted.Results: The duration needed for the RBC lysis using PL and HB was longer than that with FL (15–20 minutes vs. 10 minutes). The morphology of leukocytes was better preserved with the FL as compared to the HB and PL. The MFI of most of the antigens was similar for the HB and PL, butit was significantly lower for the FL. Conclusion: We should be aware of the changes that these agents may bring to the physical or immunophenotypic properties so as to choose the appropriate agent.en_US
dc.identifier.affiliationsAdditional Professor, Department of Hematology Sanjay Gandhi Post Graduate Institute of Medical Sciences Lucknow, U.P. Indiaen_US
dc.identifier.affiliationsB Sc MLT, Student, Dept of Hematology, SGPGIMS, Lucknow, Indiaen_US
dc.identifier.affiliationsTechnical Officer, Dept of Hematology, SGPGIMS, Lucknow, Indiaen_US
dc.identifier.affiliationsSenior Resident, Dept of Hematology, SGPGIMS, Lucknow, Indiaen_US
dc.identifier.affiliationsAssociate Professor, Deptt of BHI, SGPGIMS, Lucknow, Indiaen_US
dc.identifier.affiliationsAssistant Professor, Dept of Hematology, SGPGIMS, Lucknow, Indiaen_US
dc.identifier.affiliationsAssistant Professor, Dept of Hematology, SGPGIMS, Lucknow, Indiaen_US
dc.identifier.affiliationsAdditional Professor, Dept of Hematology, SGPGIMS, Lucknow, Indiaen_US
dc.identifier.citationRahman K, Khan MA, Sarkar MK, Biswas S, Kumar A, Singh MK, Chandra D, Gupta R.. Effect of red blood cell lysing agent in leukemia/lymphoma immunophenotyping by flow cytometry . International Archives of BioMedical and Clinical Research. 2024 Mar; 10(1): PA1-PA4en_US
dc.identifier.issn2454-9894
dc.identifier.issn2454-9886
dc.identifier.placeIndiaen_US
dc.identifier.urihttps://imsear.searo.who.int/handle/123456789/242633
dc.languageenen_US
dc.publisherMedicozum Limiteden_US
dc.relation.issuenumber1en_US
dc.relation.volume10en_US
dc.source.urihttps://doi.org/10.21276/ak4wt566en_US
dc.subjectLeukemia/Lymphoma Immunophenotypingen_US
dc.subjectRBC Lysing Agenten_US
dc.subjectLyse-Stain Wash Protocolen_US
dc.titleEffect of red blood cell lysing agent in leukemia/lymphoma immunophenotyping by flow cytometryen_US
dc.typeJournal Articleen_US
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