2010 Volume 20 Issue 2 Pages 27-33
Aim. We are in charge of the central diagnosis and cell preservation as a part of childhood acute lymphoblastic leukemia treatment study in Tokyo children’s cancer study group. It is necessary to diagnose with a minimal quantity of specimen, to preserve leukemic cells effectively as possible. On the other hand, recent progress in multi-color flow cytometry enable to analyze cell marker of leukemia in more detail. We therefore we intended to perform a immuno-phenotypic diagnosis of childhood acute lymphoblastic leukemia by nine-color analysis with digital flow cytometer.
Methods. We examined cell markers of childhood acute lymphoblastic leukemia cells by nine-color analysis using digital flow cytometers. We decided the combination of the monoclonal antibodies for nine color analysis based on the recommendation of Japan Pediatric Leukemia/Lymphoma Study Group using commercially available fluorescence-laveled antibodies.
Results. Nine colors that we used in this study were fluorescein isothyocyanate, phycoerythrin (PE), phycoerythrin-Texas Red, Peridinin Chlorophyll Protein - cyanin (Cy) 5.5, PE-Cy7, allophycocyanin (APC), APC-Cy7, Pacific Blue, and Cascade Yellow. Using list mode compensation, nine color marker analysis of childhood leukemia was easy to perform.
Discussion. Although several problems need to be solved are present, nine-color analysis using digital flow cytometer is useful to obtain precise information of antigen expression as well as save precious specimen of childhood acute lymphoblastic leukemia.
