A critical review of published methods for analysis of red cell antigen-antibody reactions by flow cytometry, and approaches for resolving problems with red cell agglutination

Transfusion Medicine Reviews
Patricia A Arndt, George Garratty

Abstract

Flow cytometry operators often apply familiar white blood cell (WBC) methods when studying red blood cell (RBC) antigens and antibodies. Some WBC methods are not appropriate for RBCs, as the analysis of RBCs requires special considerations, for example, avoidance of agglutination. One hundred seventy-six published articles from 88 groups studying RBC interactions were reviewed. Three fourths of groups used at least one unnecessary WBC procedure for RBCs, and about one fourth did not use any method to prevent/disperse RBC agglutination. Flow cytometric studies were performed to determine the effect of RBC agglutination on results and compare different methods of preventing and/or dispersing agglutination. The presence of RBC agglutinates have been shown to be affected by the type of pipette tip used for mixing RBC suspensions, the number of antigen sites/RBC, the type and concentration of primary antibody, and the type of secondary antibody. For quantitation methods, for example, fetal maternal hemorrhage, the presence of agglutinates have been shown to adversely affect results (fewer fetal D+ RBCs detected).

References

Jan 1, 1991·Vox Sanguinis·Z N BernemanM E Peetermans
Mar 1, 1989·American Journal of Clinical Pathology·S J NanceG Garratty
Apr 1, 1987·Transfusion Medicine Reviews·G Garratty
Jul 16, 1987·Journal of Immunological Methods·S J Nance, G Garratty
Apr 1, 1995·British Journal of Haematology·D BlanchardJ Y Muller
Dec 10, 1999·Cytometry·G Garratty, P A Arndt
Dec 21, 2000·Transfusion Medicine·P A Arndt, G Garratty
Jun 26, 2003·Transfusion·John D RobackChristopher D Hillyer
Feb 14, 2004·Transfusion·John D RobackChristopher D Hillyer
May 30, 2008·American Journal of Hematology·Patricia A Arndt, Belinda M Kumpel

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Citations

Oct 2, 2014·International Journal of Laboratory Hematology·M A AlzateJ M González
Sep 2, 2016·Scientific Reports·Daniela M BorgmannJaroslaw Jacak
Jan 31, 2017·Archives of Pathology & Laboratory Medicine·Victoria Parker, Christopher A Tormey

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