Relevance of sample preparation for flow cytometry

Relevance of sample preparation for flow cytometry INTRODUCTIONOver the past decades, multiparameter flow cytometry has become the method of choice for the differential diagnosis of several hematological diseases, for the definition of prognostic factors, and for the identification of rare cell populations in peripheral blood (PB) and bone marrow samples (BM).Flow cytometry, through the presence or absence of specific surface antigens, detects abnormal cells and helps in identifying their lineage and maturation stage, detects abnormal cells through identification of antigen expression that differs from the normal. The wider application of flow cytometry in diagnostic and research fields increases the need to distinguish between real and artifact positive signals given that antigen‐antibody interactions, characterized by weak bindings, may involve unexpected molecules. The presence of dead cells, cell doublets, and the nonspecific antibody binding can compete to generate artifacts in the phenotype. In some cases, an antibody can bind similar antigens not of interest or an epitope can be shared by different antigens or the Fc of many antibodies can be bound by Fc‐receptors expressed by different cells of the immune system. Undesired antibody's binding depends not only on the antibody's specificity but also on a wide variety of other interactions, as well as the physical and biological http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png International Journal of Laboratory Hematology Wiley
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Publisher
Wiley Subscription Services, Inc., A Wiley Company
Copyright
Copyright © 2018 John Wiley & Sons Ltd
ISSN
1751-5521
eISSN
1751-553X
D.O.I.
10.1111/ijlh.12755
Publisher site
See Article on Publisher Site

Abstract

INTRODUCTIONOver the past decades, multiparameter flow cytometry has become the method of choice for the differential diagnosis of several hematological diseases, for the definition of prognostic factors, and for the identification of rare cell populations in peripheral blood (PB) and bone marrow samples (BM).Flow cytometry, through the presence or absence of specific surface antigens, detects abnormal cells and helps in identifying their lineage and maturation stage, detects abnormal cells through identification of antigen expression that differs from the normal. The wider application of flow cytometry in diagnostic and research fields increases the need to distinguish between real and artifact positive signals given that antigen‐antibody interactions, characterized by weak bindings, may involve unexpected molecules. The presence of dead cells, cell doublets, and the nonspecific antibody binding can compete to generate artifacts in the phenotype. In some cases, an antibody can bind similar antigens not of interest or an epitope can be shared by different antigens or the Fc of many antibodies can be bound by Fc‐receptors expressed by different cells of the immune system. Undesired antibody's binding depends not only on the antibody's specificity but also on a wide variety of other interactions, as well as the physical and biological

Journal

International Journal of Laboratory HematologyWiley

Published: Jan 1, 2018

Keywords: ; ; ; ;

References

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