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. Therefore a diagnosis of childhood acute lymphoblastic leukemia by four-color analysis with digital flow cytometer has been examined.
Methods. We examined cell markers of childhood acute lymphoblastic leukemia cells by four-color analysis using digital flow cytometers. We selected the monoclonal antibodies for the diagnosis based on the recommendation of Japan Pediatric Lymphoma Study Group and made out a panel of antibodies which enable us to confirm aberrant antigen-expressions on the leukemic cells.
Results. Four colors that we used in this study were fluorescein isothyocyanate, phycoerythrin, phycoerythrin-cyanin 5.1, and phycoerythrin-cyanin-7. The most of childhood acute lymphoblastic leukemia cases could be diagnosed without CD45-gating. List mode compensation was useful to re-investigate specimens which was difficult to re-examine, because there were very few.
Discussion. Four-color analysis using digital flow cytometer is useful to save precious specimen of childhood acute lymphoblastic leukemia. We are intending to perform five-color analysis with CD45-gating as a next step.
