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Updated: Jan 19, 2026

Genetic Barcoding with Fluorescent Proteins for Multiplexed Applications
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Fluorescent Cell Barcoding for Immunophenotyping.

Valentina Giudice1, Giovanna Fantoni2, Angélique Biancotto3

  • 1Department of Medicine, Surgery, and Dentistry, Scuola Medica Salernitana, University of Salerno, Baronissi, Italy.

Methods in Molecular Biology (Clifton, N.J.)
|September 16, 2019
PubMed
Summary
This summary is machine-generated.

Fluorescent cell barcoding (FCB) enables multiplexing in flow cytometry, reducing technical variations and increasing data throughput. This method uniquely labels individual samples before mixing, staining, and analysis as one batch.

Keywords:
Fluorescent cell barcodingImmunophenotypingMultiplexing

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Area of Science:

  • Immunology
  • Biotechnology
  • Analytical Chemistry

Background:

  • Flow cytometry is crucial for immunophenotyping but benefits from multiplexing for robust data.
  • Multiplexing reduces antibody use, batch effects, and technical variations in flow cytometry.
  • Fluorescent cell barcoding (FCB) is a key multiplexing strategy.

Purpose of the Study:

  • To detail the protocol for fluorescent cell barcoding (FCB).
  • To provide recommendations for selecting barcoding dyes and concentrations.
  • To highlight technical considerations for implementing FCB in flow cytometry.

Main Methods:

  • FCB involves uniquely labeling individual samples with varying concentrations of fluorescent dyes.
  • Labeled samples are pooled, stained, and analyzed together as a single sample.
  • FCB facilitates normalization using a bridge control sample.

Main Results:

  • FCB significantly decreases technical variations in flow cytometry data.
  • The assay increases throughput and acquisition speed.
  • FCB simplifies the implementation of normalization strategies.

Conclusions:

  • FCB is an effective high-throughput method for multiplexing flow cytometry samples.
  • The protocol and recommendations aid in optimizing FCB implementation.
  • FCB enhances the reliability and efficiency of immunophenotyping studies.