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A method to study apoptosis in eosinophils by flow cytometry.

K Sandström¹, L Håkansson, A Lukinius

¹Department of Medical Sciences, Clinical Chemistry, Uppsala University, Sweden. kristina.sandstrom@medsci.uu.se

Journal of Immunological Methods

|June 16, 2000

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This study presents a simple flow cytometry method to distinguish between living and apoptotic eosinophils. The technique utilizes forward- and side-scatter patterns and propidium iodide staining for accurate cell discrimination.

Area of Science:

Immunology
Cell Biology
Biotechnology

Background:

Eosinophil apoptosis is crucial in regulating immune responses.
Accurate methods for studying eosinophil apoptosis are needed.

Purpose of the Study:

To develop a straightforward flow cytometry procedure for analyzing eosinophil apoptosis.
To differentiate between viable and apoptotic eosinophils in vitro.

Main Methods:

Isolation of eosinophils from healthy individuals' peripheral blood.
Culture of eosinophils and analysis of forward- and side-scatter (FS-SSC) patterns.
Staining with FDA, PI, and anti-CD95, followed by flow cytometry and cell sorting.

Main Results:

Two distinct eosinophil populations (A and B) emerged after 24 hours in culture based on FS-SSC patterns.

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Population B, characterized by lower FS scatter and PI staining, indicated hypodiploid DNA, consistent with apoptosis.

Anti-CD95 staining increased in population B over time, further confirming apoptosis.

Conclusions:

Flow cytometry, using FS-SSC patterns and PI staining, can effectively discriminate between apoptotic and living eosinophils.
The developed method provides a simple and reliable approach to study eosinophil apoptosis.