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Published on: March 25, 2016
Flow cytometric analysis of lymphomas and acute leukemias
1Department of Pathology, Norwood Clinic/Carraway Methodist Medical Center, Birmingham, Alabama 35234.
Insights
Flow cytometry is crucial for detecting and classifying leukemias and lymphomas. This technique, combined with immunophenotypic analysis, aids in disease diagnosis and subtyping based on cell characteristics.
Area of Science:
- Hematology
- Immunology
- Cell Biology
Background:
- Flow cytometry enables rapid single-cell suspension preparation, facilitating leukemia and lymphoma detection.
- Composite immunophenotypic analysis is vital for classifying leukemias and diagnosing lymphoproliferative disorders.
Purpose of the Study:
- To highlight the applicability of flow cytometry in diagnosing and classifying hematologic malignancies.
- To detail how cell size, DNA ploidy, and cell cycle analysis aid in subtyping lymphoproliferative disorders.
Main Methods:
- Utilizing flow cytometry for single-cell suspension preparation.
- Performing composite immunophenotypic analysis for disease classification.
- Analyzing cell size (light scatter), DNA ploidy, and cell cycle fractions for subtyping.
Main Results:
- Flow cytometry, with immunophenotyping, is effective for leukemia and lymphoma detection and classification.
- Specific cell characteristics (size, ploidy, synthetic fraction) correlate with lymphoma grades: low-grade (small, monoclonal, diploid, 5% synthetic fraction), intermediate (mixed sizes, variable ploidy, 5-15% synthetic fraction), and high-grade (intermediate size, diploid/near-diploid, >15% synthetic fraction).
Conclusions:
- Flow cytometry is a powerful tool for diagnosing and subtyping hematologic malignancies, particularly lymphomas.
- Accurate interpretation requires a strong understanding of disease biology and rigorous laboratory professional training.
Abstract:
In summary, flow cytometry is highly applicable to the detection and classification of leukemias and lymphomas due to the ease with which single-cell suspensions may be made. Composite immunophenotypic analysis is essential for classifying leukemias once the disease is detected by traditional means. In contrast, in the detection of lymphoproliferative diseases, the composite immunophenotype is itself diagnostic of the disease process. Characterization of size, as measured by light scatter and by DNA ploidy and cell cycle analysis, contributes to the further subdivision of lymphoproliferative disorders. Specifically, small, monoclonal cells that are diploid with a synthetic fraction of 5% are characteristic of low-grade lymphomas. Admixtures of large and small cells wherein the large cells are monoclonal and the small cells are either monoclonal or heterogeneous may be seen in intermediate lymphomas. In this category, DNA ploidy is variable and the total synthetic fraction is usually between 5% and 15%. High-grade lymphomas, with the exception of the immunoblastic category, are usually of intermediate size, are diploid or near-diploid, and exhibit synthetic fractions greater than 15%. Interestingly, few reactive T cells are seen. Ongoing efforts to standardize procedures will eventually result in more widespread applicability together with improved understanding of the attributes and limitations of this technology. The most important consideration, however, is that the technology is useless in the absence of a working knowledge of the biology of the diseases to be characterized. Conversely, the complexity of flow cytometry is sufficient to warrant rigorous training of laboratory professionals in this field.
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