Immunophenotyping of Acute Leukemia by Flow Cytometric Analysis:Use of CD45 and Right-Angle Light Scatter to Gate on Leukemic Blasts in Three-Color Analysis
| Autor: | G T Stelzer, Michael J. Borowitz, K L Guenther, K E Shults |
|---|---|
| Rok vydání: | 1993 |
| Předmět: |
Pathology
medicine.medical_specialty Myeloid Population Immunophenotyping Flow cytometry Bone Marrow Acute lymphocytic leukemia Humans Medicine education Acute leukemia education.field_of_study medicine.diagnostic_test business.industry General Medicine Precursor Cell Lymphoblastic Leukemia-Lymphoma Flow Cytometry medicine.disease Molecular biology Leukemia Myeloid Acute Leukemia medicine.anatomical_structure Leukocyte Common Antigens Bone marrow business |
| Zdroj: | American Journal of Clinical Pathology. 100:534-540 |
| ISSN: | 1943-7722 0002-9173 |
| DOI: | 10.1093/ajcp/100.5.534 |
| Popis: | This article describes a procedure for performing routine three-color flow cytometric analysis for acute leukemia on lysed whole bone marrow preparations. This technique uses the combination of CD45 intensity and right-angle light scatter (RALS) to distinguish leukemic cells from normal lymphocytes, monocytes, neutrophils, eosinophils, and nucleated red blood cells. On this display, leukemic cells occupy a unique blast region characterized by intermediate CD45 density and low RALS, which, in normal marrows, contains less than 5% of the total cells. This approach was applied to 39 cases of acute leukemia and 8 cases of myelodysplasia or myeloproliferative disorders. The estimate of blasts by flow cytometric analysis was correlated highly with morphologic leukemic cell counts over a wide range. Moreover, the pattern seen on the CD45-RALS display was different for different French-American-British subtypes of leukemia, suggesting that this pattern might be useful for categorization. When CD45-peridin chlorophyll alpha protein was combined with other pairs of fluorescein isothiocyanate- and phycoerythrin-conjugated reagents, it was possible to set an analysis window on the leukemic blasts and display dual-parameter (ie, green vs. red fluorescence) data regarding expression of two additional markers on the leukemic population. This gating strategy was superior to traditional forward-angle versus RALS displays in that it did a better job of isolating the leukemic cells analytically. |
| Databáze: | OpenAIRE |
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