Related Experiment Video
Updated: Aug 8, 2026
Flow Cytometry to Estimate Leukemia Stem Cells in Primary Acute Myeloid Leukemia and in Patient-derived-xenografts, at Diagnosis and Follow Up
Published on: March 26, 2018
[Immunophenotyping of eighty six children with acute lymphoblastic leukemia by three-color flow cytometry]
Suigui Wan1, Wenyu Gong, Xuejing Sun
1Department of Hematology, Xuanwu Hospital, CUMS, Beijing 100053, China.
Insights
Three-color flow cytometry with CD(45)/SSC gating improves specificity in childhood acute lymphoblastic leukemia (ALL) immunophenotyping. This method reduces false positives, enhancing diagnostic reliability for B-ALL and T-ALL subtypes.
Area of Science:
- Hematology
- Immunology
- Oncology
Context:
- Childhood acute lymphoblastic leukemia (ALL) diagnosis relies heavily on accurate immunophenotyping.
- Traditional methods may face challenges with specificity and distinguishing malignant from normal cells.
Purpose:
- To assess the specificity of three-color flow cytometry using CD(45)/SSC gating for childhood ALL immunophenotyping.
- To evaluate the reliability and accuracy of this technique in classifying ALL subtypes.
Summary:
- Three-color flow cytometry with CD(45)/SSC gating demonstrated high specificity in childhood ALL immunophenotyping.
- This method showed a better correlation with leukemic cell count than FSC/SSC gating, with low false positive rates.
- Analysis of 86 ALL cases revealed 95.3% B-ALL (76.8% common-ALL, 6.1% Pro-B-ALL) and 2.3% T-ALL. CD(34)(+) and myeloid-associated antigen expression were noted in 57.0% and 34.9% of cases, respectively.
Impact:
- CD(45)/SSC gating effectively eliminates interference from normal bone marrow cells, leading to more reliable immunophenotyping results.
- This enhanced specificity aids in precise diagnosis and classification of childhood ALL.
- Improved diagnostic accuracy can inform treatment strategies and prognosis for pediatric leukemia patients.
Objectives:
To evaluate the specificity of three-color flow cytometry in childhood acute lymphoblastic leukemia (ALL) immunophenotyping.
Methods:
Immunophenotyping was performed by three-color flow cytometry analysis using CD(45)/SSC gating.
Results:
The percentage of blasts was correlated better with leukemic cell count compared with that of FSC/SSC, and the false positive results were low. Among eighty six cases of ALL, 95.3% was B-ALL, in which common-ALL and Pro-B-ALL were 76.8% and 6.1%, respectively, and 2.3% was T-ALL. CD(34)(+) and myeloid-associated antigen expression were observed in 57.0% and 34.9% of the cases, respectively, among which Pro-B-ALL was the commonest. CD(33) was more commonly expressed than CD(13) in Pro-B-ALL cases, but no difference in the expression between these two antigens in other subtypes.
Conclusion:
Gating of CD(45)/SSC eliminated effection of normal cells to blasts in bone marrow, with which the immunophenotyping results were more reliable.
