open access

Vol 48, No 3 (2017)
Prace poglądowe / Reviews
Published online: 2017-07-01
Submitted: 2017-06-28
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Application of flow cytometry to immunohematological tests of red blood cells

Jadwiga Fabijańska-Mitek, Anna Stachurska1
DOI: 10.1016/j.achaem.2017.07.005
·
Acta Haematol Pol 2017;48(3):183-188.
Affiliations
  1. Zakład Immunohematologii, Centrum Medyczne Kształcenia Podyplomowego, Warszawa, Polska

open access

Vol 48, No 3 (2017)
Prace poglądowe / Reviews
Published online: 2017-07-01
Submitted: 2017-06-28

Abstract

Flow cytometry (FC) has been primarily applied to the diagnosis of hematological malignancies, and thereafter, to detection and quantification of CD34+ cells in bone marrow transplants, and granulocytes in neutropenias and paroxysmal nocturnal hemoglobinuria (PNH). In PNH and hereditary spherocytosis, changes in some of the erythrocyte membrane proteins are tested (CD59, CD55, and band 3). The purpose of this paper is to focus on the use of FC in RBC testing. With anti-D, -HbF, and -CA (carbonic anhydrase), we can detect RhD+, HbF+, and CA- fetal RBCs in the maternal RhD-, HbF-, and CA+ blood sample. Obtained results allow to select the appropriate dose of anti-D Ig in the RhD prophylaxis of feto-maternal incompatibility, or to detect the cause of fetal anemia. Expression of antigens and their weak variants, and concentration of specific antibodies, can also be assessed. It is possible to observe changes in selected CD molecules during storage of RBC units. If RBCs for transfusion are unavailable, due to patient's unusual antibody specificity, some of the available RBCs are opsonised, and then phagocytosis with the recipient monocytes is assessed. The microscopic time-consuming and subjective assay is usually used. Stained CD14+ monocytes and CD235a+ erythrocytes are visible on cytograms as well as their interaction. It makes evaluation of phagocytosis easier and objective. Microparticles of RBCs released during storage are also detected. They are 235a+. In differentiation of hemolysis causes, it is important to measure osmotic fragility, and that can be also achieved using FC. Flow cytometry should be applied to immunohematological testing of red blood cells more often than now.

Abstract

Flow cytometry (FC) has been primarily applied to the diagnosis of hematological malignancies, and thereafter, to detection and quantification of CD34+ cells in bone marrow transplants, and granulocytes in neutropenias and paroxysmal nocturnal hemoglobinuria (PNH). In PNH and hereditary spherocytosis, changes in some of the erythrocyte membrane proteins are tested (CD59, CD55, and band 3). The purpose of this paper is to focus on the use of FC in RBC testing. With anti-D, -HbF, and -CA (carbonic anhydrase), we can detect RhD+, HbF+, and CA- fetal RBCs in the maternal RhD-, HbF-, and CA+ blood sample. Obtained results allow to select the appropriate dose of anti-D Ig in the RhD prophylaxis of feto-maternal incompatibility, or to detect the cause of fetal anemia. Expression of antigens and their weak variants, and concentration of specific antibodies, can also be assessed. It is possible to observe changes in selected CD molecules during storage of RBC units. If RBCs for transfusion are unavailable, due to patient's unusual antibody specificity, some of the available RBCs are opsonised, and then phagocytosis with the recipient monocytes is assessed. The microscopic time-consuming and subjective assay is usually used. Stained CD14+ monocytes and CD235a+ erythrocytes are visible on cytograms as well as their interaction. It makes evaluation of phagocytosis easier and objective. Microparticles of RBCs released during storage are also detected. They are 235a+. In differentiation of hemolysis causes, it is important to measure osmotic fragility, and that can be also achieved using FC. Flow cytometry should be applied to immunohematological testing of red blood cells more often than now.

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Keywords

Flow cytometry; Feto-maternal hemorrhage; Erythrophagocytosis; CD of erythrocytes; RBCs storage; Microparticles

About this article
Title

Application of flow cytometry to immunohematological tests of red blood cells

Journal

Acta Haematologica Polonica

Issue

Vol 48, No 3 (2017)

Pages

183-188

Published online

2017-07-01

DOI

10.1016/j.achaem.2017.07.005

Bibliographic record

Acta Haematol Pol 2017;48(3):183-188.

Keywords

Flow cytometry
Feto-maternal hemorrhage
Erythrophagocytosis
CD of erythrocytes
RBCs storage
Microparticles

Authors

Jadwiga Fabijańska-Mitek
Anna Stachurska

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