THE USE OF INDIRECT IMMUNOFLUORESCENCE FLOW CYTOMETORY FOR THE ANALYSIS OF ERYTHROCYTE ANTIGEN (I)

• Published in: Journal of the Japan Society of Blood Transfusion Vol. 33; no. 6; pp. 695 - 701
• Main Authors: Okubo, Y., Seno, Taiko, Tanaka, Mitsunobu, Yamaguchi, H.
• Format: Journal Article
• Language: Japanese
• Published: The Japan Society of Transfusion Medicine and Cell Therapy 1987
• ISSN: 0546-1448; 1883-8383
• DOI: 10.3925/jjtc1958.33.695

Zygosity determinations are important in population and family studies. The purpose of this study is to determine whether an Rh phenotype of red cells is derived from a homozugous or heterozygous subject by the use of the indirect immunoflurorescence Flow Cytometry (SPIII) without family studies. As a preliminary study, conditions for the sensitization of red cells with antibodies were investigated. We found purchesed anti-Rh typing sera and mouse monoclonal antibodies prepared in our labortory to be usable in the 8-to 32-fold dilutions. The best condition for the primary sensitization of red cells with the sera mentioned above was to incubate them at 37°C for 1 hour and overnight at 4°C. The secondary sensitization was carried out with 100-fold diluted FITC labeled antihuman or antimouse globulin at 4°C for 30 mintes. In this sutdy selection of the suitable Rh-antibody was most important.