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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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"Immuno-FlowFISH": Applications for Chronic Lymphocytic Leukemia.

Henry Y L Hui1, Wendy N Erber1,2, Kathy A Fuller3

  • 1Translational Cancer Pathology Laboratory, School of Biomedical Sciences (M504), The University of Western Australia, Crawley, WA, Australia.

Methods in Molecular Biology (Clifton, N.J.)
|April 19, 2023
PubMed
Summary

A new Immuno-flowFISH method enhances leukemia diagnosis by integrating imaging flow cytometry and FISH. This approach offers superior accuracy for detecting chromosomal abnormalities in Chronic Lymphocytic Leukemia (CLL) cells.

Keywords:
Chronic lymphocytic leukemiaFluorescence in situ hybridizationImaging flow cytometry

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Area of Science:

  • Hematology
  • Cellular Biology
  • Genetics

Background:

  • Current diagnostic methods for hematological malignancies have limitations in detecting phenotypic and genetic changes.
  • Bridging the gap between clinical assessment and molecular diagnostics is crucial for accurate disease evaluation.

Purpose of the Study:

  • To develop and optimize an integrated method, Immuno-flowFISH, for advanced single-cell analysis.
  • To leverage imaging flow cytometry for enhanced detection of chromosomal abnormalities in Chronic Lymphocytic Leukemia (CLL).

Main Methods:

  • Developed the "Immuno-flowFISH" technique combining imaging flow cytometry and fluorescence in situ hybridization (FISH).
  • Optimized the protocol for detecting specific chromosomal abnormalities (trisomy 12 and del(17p)) in CD19/CD5+ CD3- CLL cells.
  • Established a detailed workflow with quality control considerations for CLL analysis.

Main Results:

  • The Immuno-flowFISH method accurately detects numerical and structural chromosomal abnormalities in CLL cells.
  • This integrated methodology demonstrates greater accuracy and precision compared to standard FISH.
  • The protocol enables simultaneous assessment of phenotypic and genetic markers at the single-cell level.

Conclusions:

  • Immuno-flowFISH represents a next-generation protocol for the holistic cellular assessment of hematological disorders.
  • This method offers advancements for both research and clinical laboratory settings in disease analysis.
  • The technique provides a more comprehensive understanding of disease at the single-cell level.