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Updated: Jul 11, 2026
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Discrimination of Seven Immune Cell Subsets by Two-fluorochrome Flow Cytometry
Published on: March 5, 2019
"6 markers/5 colors" extended white blood cell differential by flow cytometry
Jean-Luc Faucher1, Charlotte Lacronique-Gazaille, Elise Frébet
1Laboratoire d'Hématologie, Hôpital Dupuytren, Limoges, France.
Summary
Flow cytometry (FCM) offers a more comprehensive white blood cell (WBC) differential than standard cytology. FCM accurately quantifies cell types and identifies abnormalities, including blast cells and inflammatory monocytes.
Area of Science:
- Hematology
- Immunology
- Analytical Chemistry
Background:
- Standard white blood cell (WBC) differential using cytology is limited to five cell types and qualitative abnormal cell identification.
- Current methods are often problematic, poorly reproducible, and labor-intensive.
Purpose of the Study:
- To evaluate the efficacy of flow cytometry (FCM) for WBC differential analysis.
- To compare FCM's performance against standard cytology for detecting and quantifying various cell types and abnormalities.
Main Methods:
- Utilized FCM with a 6-marker, 5-color panel (CD36-FITC/CD2-PE+CRTH2-PE/CD19-ECD/CD16-Cy5/CD45-Cy7) on 379 subjects.
- Detected and quantified 12 circulating cell types, with 11 being quantified.
Main Results:
- FCM demonstrated comparable sensitivity and specificity to standard cytology for quantifying WBCs, neutrophils, eosinophils, basophils, monocytes, and lymphocytes.
- FCM excelled in detecting and quantifying blast cells and immature granulocytes, providing lineage orientation.
- FCM identified lymphocytosis with lineage assignment and detected inflammatory syndromes by identifying CD16pos monocytes.
Conclusions:
- WBC differential by FCM is as reliable as standard cytology.
- FCM offers superior identification and quantification of parameters not assessable by standard cytology, such as blast cell lineage and inflammatory monocyte markers.
