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The development of flow cytometry techniques began in 1934 with initial attempts by Andrew Moldavan, a bacteriologist who counted the cells in a flowing capillary system. Moldavan pumped cells through a capillary tube focused under a microscope for visualization. The invention of photometry allowed the measurement of differentially-stained cells, and Louis Kamentsky developed the first multiparameter flow cytometer in 1965 to identify and count the cancer cells in cervical tissue specimens.
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Related Experiment Video

Updated: Jan 21, 2026

Flow Cytometry to Estimate Leukemia Stem Cells in Primary Acute Myeloid Leukemia and in Patient-derived-xenografts, at Diagnosis and Follow Up
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Multiparametric Flow Cytometry in Mixed Phenotype Acute Leukemia.

Sindhura Lakshmi Koulmane Laxminarayana1, Nishika Madireddy2, Chethan Manohar1

  • 11Department of Pathology, Kasturba Medical College, Manipal Academy of Higher Education, Manipal, Udupi, Karnataka 576104 India.

Indian Journal of Hematology & Blood Transfusion : an Official Journal of Indian Society of Hematology and Blood Transfusion
|August 8, 2019
PubMed
Summary

Mixed phenotype acute leukaemia (MPAL) is rare, affecting 4.1% of acute leukaemia cases. Accurate diagnosis using multiparametric flow cytometry (FCM) is crucial for appropriate treatment and improved outcomes in this ambiguous lineage leukemia.

Keywords:
ImmunophenotypeLeukemia of ambiguous lineageMixed phenotype acute leukemiaMultiparametric flow cytometry

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Area of Science:

  • Hematology
  • Oncology
  • Immunology

Background:

  • Mixed phenotype acute leukaemia (MPAL) is a rare and complex leukemia characterized by blasts expressing antigens from more than one lineage.
  • Accurate diagnosis is challenging due to the ambiguous lineage of the blasts.
  • Current diagnostic criteria include the European Group for the Immunological Classification of Leukaemias (EGIL) and World Health Organization (WHO) guidelines.

Purpose of the Study:

  • To determine the incidence of MPAL in a cohort of acute leukaemia cases.
  • To analyze the immunophenotypic characteristics, genetic abnormalities, and treatment outcomes of MPAL patients.
  • To emphasize the importance of multiparametric flow cytometry (FCM) for accurate MPAL diagnosis.

Main Methods:

  • Retrospective analysis of 218 consecutive acute leukaemia cases diagnosed using multiparametric FCM.
  • MPAL cases were identified based on EGIL and WHO 2008/2016 criteria.
  • Immunophenotypic data, cytogenetics, and treatment responses were reviewed.

Main Results:

  • MPAL accounted for 4.1% (9/218) of acute leukaemia cases.
  • The majority of MPAL patients were male (88.8%) and younger than 20 years (44.4%).
  • Common MPAL subtypes included B/T, T/myeloid, and B/myeloid.
  • Philadelphia chromosome positivity was observed in B/myeloid MPAL and CML in B/myeloid blast crisis.
  • Treatment with acute lymphoblastic leukaemia regimens resulted in complete remission in 2 out of 7 patients.
  • High early mortality (62.5%) due to febrile neutropenia was noted.

Conclusions:

  • Multiparametric FCM is essential for the accurate and early diagnosis of MPAL.
  • Limited antibody panels may lead to misdiagnosis of MPAL.
  • MPAL presents unique diagnostic and therapeutic challenges, with significant early mortality.
  • Further research into optimized treatment strategies for MPAL is warranted.