Related Experiment Videos
[Flow cytometry in clinical laboratory medicine]
K Ohmori1, T Yoneda, S Sugiyama
1Department of Laboratory Medicine, Kyoto University, School of Medicine.
Insights
Accurate lymphocyte gating in flow cytometry is crucial for diagnosing blood disorders and leukemia. Utilizing CD13 and CD33 markers improves precision in identifying lymphocyte subsets and monitoring non-lymphoid cell contamination.
Area of Science:
- Hematology
- Immunology
- Cell Biology
Context:
- Flow cytometry is essential for evaluating immunological functions and diagnosing leukemias.
- Accurate gating of lymphocyte subsets is critical for reliable analysis.
- Current methods may require optimization for specific patient populations like those with paroxysmal nocturnal hemoglobinuria (PNH).
Purpose:
- To recommend improved gating strategies for lymphocyte subset analysis in flow cytometry.
- To enhance the accuracy of differential diagnosis for leukemias.
- To highlight the utility of novel cell adhesion molecules in leukemia research.
Summary:
- The study proposes using CD13 and CD33, in addition to CD45 and CD14, for more precise lymphocyte gating and monitoring of non-lymphoid cell contamination, particularly in PNH patients.
- It advocates for increased use of 2-color and 3-color analyses in routine lymphocyte subset testing.
- Combining nuclear DNA content or TdT with other markers aids in analyzing minimal residual disease in leukemia.
Impact:
- Improved diagnostic accuracy for various blood disorders and leukemias.
- Enhanced monitoring of minimal residual disease in leukemia patients.
- Provides insights into the role of cell adhesion molecules in leukocyte recruitment and leukemia cell infiltration.
Abstract:
Flow cytometric analysis of blood cells is an important technique for the evaluation of the immunological functions of patients with various disorders, and also for the differential diagnosis of leukemias. In the lymphocyte subset analysis, special attention should be paid to the optimal gating of lymphocytes. We recommend the use of CD13 and CD33 to ascertain more accurate gating of lymphocytes and more efficient monitoring of the contamination of non-lymphoid cells, especially in patients with PNH, in addition to CD45 and CD14, which are already utilized routinely. We also feel that 2-color and 3-color analyses should be applied more frequently for the routine laboratory tests for lymphocyte subsets. With regard to the analysis of leukemia cells, the combined use of nuclear DNA content or TdT (terminal deoxynucleotidyl transferase) with other routine cellular markers is quite useful for the analysis of a small number of leukemia cells, and enables the differentiation of leukemia cells from the contaminating non-malignant cells, particularly in the analysis of the minimal residual diseases. Among the cellular markers defined by specific monoclonal antibodies, we stress the physiological significance of recently developed cell adhesion molecules, such as selectins and their carbohydrate ligands, which are closely involved in the recruitment of leukocytes in inflammatory responses, as well as in the infiltration processes of leukemia cells.