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¹The Haddow Laboratories, Institute of Cancer Research, Sutton, Surrey, UK.

Methods in Molecular Biology (Clifton, N.J.)

|February 22, 2012

Summary

This summary is machine-generated.

The introduction of phycoerythrin (PE) revolutionized multi-antigen flow cytometry. This natural fluorochrome allows simultaneous excitation with fluorescein, simplifying multicolor analysis.

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Area of Science:

Biotechnology
Immunology
Analytical Chemistry

Background:

Multi-antigen detection in flow cytometry previously required two lasers.
Excitation of fluorescein and rhodamine derivatives necessitated distinct laser sources.
This limitation complicated simultaneous antigen measurement.

Purpose of the Study:

To introduce phycoerythrin (PE) as a novel fluorochrome for flow cytometry.
To demonstrate PE's utility in simplifying multicolor analysis.
To address the need for more efficient multi-antigen detection.

Main Methods:

Utilized phycoerythrin (PE), a phycobiliprotein from red algae.
Excited PE at 488 nm, simultaneously with fluorescein.
Analyzed emission spectra and spectral overlap.

Main Results:

PE can be efficiently excited at 488 nm alongside fluorescein.
PE exhibits peak fluorescence at 578 nm, distinct from fluorescein's 520 nm peak.
Spectral overlap between PE and fluorescein requires correction.

Conclusions:

Phycoerythrin (PE) significantly advanced multicolor flow cytometry.
Simultaneous excitation of PE and fluorescein simplifies multi-antigen analysis.
Electronic or software-based spectral correction is necessary for accurate results.