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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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Updated: May 15, 2026

A Simple Flow Cytometric Method to Measure Glucose Uptake and Glucose Transporter Expression for Monocyte Subpopulations in Whole Blood

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Monochromatic gating method by flow cytometry for high purity monocyte analysis.

Linda Wong¹, Bruce H Davis

¹Trillium Diagnostics, LLC, Brewer, Maine 04412, USA.

Cytometry. Part B, Clinical Cytometry

|January 4, 2013

Summary

This summary is machine-generated.

High purity monocyte gating for quantitative flow cytometry assays is improved using a monochromatic CD45 and CD64 antibody combination. This method enhances accuracy for antigen expression analysis in myeloid cells, particularly monocytes.

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Area of Science:

Immunology
Cell Biology
Flow Cytometry

Background:

Accurate antigen expression assays on myeloid cells require high purity leukocyte subpopulation gating.
Standard CD45/side scatter (SSC) gating is insufficient for quantitative analysis of antigen changes in monocytes.
A novel monochromatic gating approach using CD45 and CD64 has been validated for high purity monocyte identification.

Purpose of the Study:

To validate a monochromatic gating strategy for high purity monocyte identification in flow cytometry.
To compare the efficacy of monochromatic CD45/CD64 gating with traditional methods for monocyte quantification.
To establish a more accurate method for quantitative antigen expression analysis on monocytes.

Main Methods:

Blood samples were stained with various antibody combinations, including monochromatic CD45 FITC + CD64 FITC.
Comparative analysis of monocyte percentages was performed using CD45/SSC gating versus CD45/CD64 and CD45/CD163 gating.
Data analysis focused on the percentage of monocytes identified by different gating strategies.

Main Results:

Monocyte recovery using monochromatic CD64 gating showed no statistical difference compared to two-color CD45 + CD64 or CD45 + CD163 gating.
All tested immunologic monocyte identification methods resulted in a 12.93%-15.15% reduction in monocyte percentage compared to CD45/SSC gating.
The validated monochromatic gating approach provides higher purity for monocyte identification.

Conclusions:

A monochromatic combination of CD45 and CD64 antibodies offers superior monocyte purity in flow cytometry compared to CD45/SSC gating.
This method enables the development of high-resolution, four-color assays for simultaneous analysis of multiple cell types and markers.
Potential applications include the detection of paroxysmal nocturnal hemoglobinuria by analyzing phosphatidylinositol (PI) linked protein expression on monocytes and neutrophils.