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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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Analysis of Side Population in Solid Tumor Cell Lines

Analysis of Side Population in Solid Tumor Cell Lines

Published on: February 23, 2021

Flow cytometry of the side population (SP).

¹Institut de Recerca Hospital Universitari Vall d'Hebron, Barcelona, Spain.

Current Protocols in Cytometry

|September 5, 2008

Summary

The side population (SP) technique aids in identifying and isolating stem cells using Hoechst 33342 dye. This guide simplifies the complex flow cytometry protocol for researchers, improving stem cell detection.

Area of Science:

Cell Biology
Stem Cell Research
Biotechnology

Background:

Side population (SP) cells are crucial for defining stem cell compartments.
SP cells are identified by their ability to efflux Hoechst 33342 (Ho342) dye.
The SP method is vital for stem cell detection and isolation via fluorescence-activated cell sorting (FACS).

Purpose of the Study:

To provide a simplified and optimized protocol for identifying and isolating side population (SP) stem cells.
To address the technical challenges associated with the Hoechst 33342 efflux assay.
To enhance the accessibility and accuracy of SP cell analysis in flow cytometry.

Main Methods:

Utilizing fluorescence-activated cell sorting (FACS) for cell analysis.
Employing Hoechst 33342 (Ho342) dye for differential dye retention assays.

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Effective Detection of Hoechst Side Population Cells by Flow Cytometry

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Isolation of the Side Population in Myc-induced T-cell Acute Lymphoblastic Leukemia in Zebrafish

Isolation of the Side Population in Myc-induced T-cell Acute Lymphoblastic Leukemia in Zebrafish

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Last Updated: Jul 2, 2026

Analysis of Side Population in Solid Tumor Cell Lines

Analysis of Side Population in Solid Tumor Cell Lines

Published on: February 23, 2021

Effective Detection of Hoechst Side Population Cells by Flow Cytometry

Effective Detection of Hoechst Side Population Cells by Flow Cytometry

Published on: August 23, 2024

Isolation of the Side Population in Myc-induced T-cell Acute Lymphoblastic Leukemia in Zebrafish

Isolation of the Side Population in Myc-induced T-cell Acute Lymphoblastic Leukemia in Zebrafish

Published on: May 4, 2017

Detailed guidance on experimental setup, acquisition conditions, and data analysis for SP detection.

Main Results:

The study details a refined protocol for SP cell identification.
The optimized method facilitates easier discrimination of SP cells based on Ho342 retention.
Improved data analysis strategies are presented for accurate SP event detection.

Conclusions:

The side population technique, while powerful, presents significant technical hurdles.
This work offers practical solutions and expert advice to overcome challenges in SP cell analysis.
The enhanced protocol aims to improve the reproducibility and success rate of SP stem cell isolation.