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Isolation of human basophils by flow microfluorometry.

G J Weil, W M Leiserson, T M Chused

Journal of Immunological Methods

|March 25, 1983

Summary

Researchers developed a cell sorting method to isolate pure basophils, a type of white blood cell, for further study. This technique achieved high purity and viability, yielding 15% of the starting cells.

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Area of Science:

Immunology
Cell Biology
Hematology

Background:

Basophils are critical white blood cells involved in allergic responses and immune regulation.
Isolating pure basophils is essential for detailed functional and molecular studies.
Previous methods for basophil isolation had limitations in purity and yield.

Purpose of the Study:

To develop and validate a high-purity cell sorting method for human basophils.
To enable downstream analysis of purified basophil populations.

Main Methods:

Human peripheral blood mononuclear cells were labeled with fluorescein-conjugated anti-IgE antibody.
A two-step cell sorting procedure was employed to enrich basophils.
Flow microfluorometric analysis was used for cell characterization and purity assessment.

Main Results:

Basophils were successfully enriched to 97-99% purity.
The viability of sorted basophils, assessed by trypan blue exclusion, exceeded 90%.
The overall net yield of the cell sorting procedure was 15%.

Conclusions:

A robust and efficient two-step cell sorting protocol can yield highly pure and viable human basophils.
This method facilitates further research into basophil function and their role in immune responses.
The developed technique is suitable for preparing basophil populations for advanced analyses.

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