Related Experiment Video
Updated: May 12, 2026
Immunophenotyping and Cell Sorting of Human MKs from Human Primary Sources or Differentiated In Vitro from Hematopoietic Progenitors
Published on: August 7, 2021
Immunophenotyping of selected hematologic disorders--focus on lymphoproliferative disorders with more than one
1Department of Pathobiology and Laboratory Medicine, University of Toronto, University Health Network, Toronto, ON, Canada. anna.porwit@uhn.ca
Insights
This study introduces cost-effective 10-color flow cytometry panels for diagnosing lymphoproliferative disorders (LPDs). These panels efficiently identify aberrant B- and T-cell populations, aiding in LPD diagnosis within budget constraints.
Area of Science:
- Clinical Immunology
- Hematology
- Laboratory Medicine
Background:
- Clinical laboratories face rising demand for flow cytometry testing amid funding limitations.
- There is a need for economical yet comprehensive immunophenotyping panels for diagnosing lymphoproliferative disorders (LPDs).
Purpose of the Study:
- To present cost-effective 10-color flow cytometry panels for the diagnosis of LPDs.
- To detail B-cell and T-cell panels that meet European Leukemia Net recommendations.
- To highlight the utility of multicolor analysis in detecting multiple aberrant cell populations in LPDs.
Main Methods:
- Application of two 10-color flow cytometry tubes for surface marker analysis in LPD diagnosis.
- Utilizing specific antibody combinations for B-cell (including plasma cell) and T/NK-cell panels.
- Employing multicolor analysis to simultaneously study multiple antigen expressions on defined cell populations.
Main Results:
- The B-cell panel includes mandatory markers for LPD diagnosis, while a separate panel targets plasma cell aberrations.
- The T-cell panel enumerates various T- and NK-cell subsets.
- Multicolor analysis effectively detects multiple aberrant populations, which occur in approximately 5% of B-cell chronic LPDs.
Conclusions:
- The developed 10-color flow cytometry panels provide sufficient immunophenotyping information for LPD diagnosis cost-effectively.
- Multicolor flow cytometry is crucial for identifying complex aberrant immunophenotypes in LPDs.
- These panels offer a practical solution for clinical laboratories managing increased testing demands and limited resources.
Abstract:
Currently, clinical laboratories face increasing demand for flow cytometry testing combined with limited funding. Therefore, many laboratories search for panels that would provide sufficient immunophenotyping information and meet economical requirements. At the Flow Cytometry Laboratory, University Health Network, Toronto, ON, Canada, we apply two 10-color tubes of surface markers for diagnosis of lymphoproliferative disorders (LPDs). These tubes contain most of the mandatory B- and T-cell markers according to European Leukemia Net (www.leukemia-net.org) recommendations. The B-cell-oriented panel includes the following antibodies: Kappa-FITC/lambda-PE/CD19-ECD/CD38-PC5.5/CD20-PC7/CD34-APC/CD23 APC-AF700/CD10 APC-AF750/CD5-PB/CD45-KO. A different combination is applied to detect cytoplasmic Ig light chain expression and aberrant immunophenotype of plasma cells. The T-cell panel allows enumeration of various T- and NK-cell subsets: CD57-FITC/CD11c-PE/CD8-ECD/CD3-PC5.5/CD2-PC7/CD56-APC/CD7-APC-AF700/CD4-APC-AF750/CD5-PB/CD45-KO. The reported overall incidence of B-cell chronic LPDs presenting with more than one aberrant population is approximately 5%. Multicolor analysis facilitates the detection of multiple aberrant populations in the same sample because expression of multiple antigens can be studied simultaneously in each defined population. Examples of LPDs with multiple aberrant populations are presented.
