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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
[Determination of intracellular molecules in cells in hematopoietic malignancy using flow cytometry]
1Ustav klinické imunologie a alergologie LF UK a FN, Hradec Králové.
Insights
A new protocol enables routine detection of intracellular molecules in hematopoetic malignancies. This method improves immunophenotypisation for diagnosing complex blood cancers when surface markers are unclear.
Area of Science:
- Hematology
- Immunology
- Cell Biology
Context:
- Immunophenotypisation is crucial for diagnosing hematopoetic malignancies.
- Detecting intracellular molecules is vital when surface marker expression is poor or results are equivocal.
Purpose:
- To establish a simple, rapid, and routine procedure for detecting intracellular molecules in hematopoetic cells.
- To enhance diagnostic accuracy for complex hematopoetic malignancies.
Summary:
- A protocol using paraformaldehyde fixation and a specific lysing solution (Lysing solution G) allows for effective permeabilisation.
- Cells are subsequently washed with a Tween 20 solution, enabling detection of nuclear and cytoplasmic molecules.
- This method is readily available for routine laboratory use.
Impact:
- Improves the diagnostic capabilities for hematopoetic malignancies, especially in challenging cases.
- Facilitates accurate diagnosis of leukemias with ectopic expression or biphenotypic characteristics.
- Provides a valuable tool for routine laboratory testing in hematopathology.
Background:
Detection of membrane molecules by immunophenotypisation is a routine counterpart of the laboratory tests which are needed for the complex diagnostic procedures of hematopoetic malignancies at present. The immunochemical detection of intracellular molecules is essential in such cases, where the expression of surface molecules on malignant cells is poor, or where the results of other diagnostic analyses are unequivocal (cases with ectopic expression of other lineage-specific molecules and cases of biphenotypic leukemias).
Methods And Results:
We followed the procedure using 4% solution of paraformaldehyd in phosphate buffered saline for fixation of cells. Lysing solution G (Becton-Dickinson) was used for permeabilisation of cells to detect nuclear and cytoplasmatic molecules. The permeabilised cell suspension was than washed with 0.5% solution of Tween 20 in phosphate buffered saline. This simple and rapid procedure is readily available for the routine detection of intracellular molecules. This approach is important for the diagnosis of hematopoetic malignancies.
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