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Flow Cytometry

Flow Cytometry

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: Jul 17, 2026

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

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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Diagnosing PNH with FLAER and multiparameter flow cytometry.

D Robert Sutherland¹, Nancy Kuek, Jeff Davidson

¹Clinical Flow Cytometry Laboratory, University Health Network, Toronto, Canada. rob.southerland@utoronto.ca

Cytometry. Part B, Clinical Cytometry

|February 8, 2007

Summary

A new FLAER assay improves diagnosis of paroxysmal nocturnal hemoglobinuria (PNH) by detecting minor PNH clones. This flow cytometry method enhances sensitivity for PNH detection in various hematologic disorders.

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Area of Science:

Hematology
Immunology
Cell Biology

Background:

Paroxysmal nocturnal hemoglobinuria (PNH) is an acquired stem cell disorder characterized by deficiency of GPI-linked proteins.
Diagnosis traditionally involves flow cytometry detecting loss of GPI-linked antigens on blood cells.
Existing flow assays have limitations, prompting the development of more sensitive methods.

Purpose of the Study:

To develop and validate a FLAER-based flow cytometry assay for diagnosing and monitoring PNH.
To enhance the detection of PNH clones, especially minor populations, in patients with other hematologic conditions.

Main Methods:

A single-tube assay was developed combining FLAER with CD45, CD33, and CD14.
This allowed simultaneous analysis of FLAER and GPI-linked CD14 on neutrophil and monocyte lineages.
Assay performance was compared to standard CD55 and CD59 analysis.

Main Results:

The FLAER assay demonstrated excellent agreement with standard CD55 and CD59 analysis.
Increased sensitivity was observed due to a higher signal-to-noise ratio, enabling detection of PNH clones as small as 1% in aplastic anemia.
Abnormal FLAER staining was also noted in blast populations in acute leukemia, suggesting broader utility.

Conclusions:

FLAER combined with multiparameter flow cytometry provides an improved method for PNH diagnosis and monitoring.
The assay shows potential for detecting unsuspected myeloproliferative disorders and minor PNH clones.