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Updated: Sep 22, 2025

Genetic Barcoding with Fluorescent Proteins for Multiplexed Applications

Genetic Barcoding with Fluorescent Proteins for Multiplexed Applications

Published on: April 14, 2015

Visual barcodes for clonal-multiplexing of live microscopy-based assays.

Tom Kaufman¹, Erez Nitzan¹, Nir Firestein¹

¹Department of Molecular Cell Biology, Weizmann Institute of Science, Rehovot, Israel.

Nature Communications

|May 18, 2022

Summary

This summary is machine-generated.

Researchers developed visual barcodes for live cell imaging, enabling simultaneous monitoring of 12 signaling pathways. This breakthrough in multiplexing advances high-content analysis for complex biological systems and cancer research.

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Last Updated: Sep 22, 2025

Genetic Barcoding with Fluorescent Proteins for Multiplexed Applications

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Published on: April 14, 2015

Multiplexed Barcoding Image Analysis for Immunoprofiling and Spatial Mapping Characterization in the Single-Cell Analysis of Paraffin Tissue Samples

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Phospho Flow Cytometry with Fluorescent Cell Barcoding for Single Cell Signaling Analysis and Biomarker Discovery

Phospho Flow Cytometry with Fluorescent Cell Barcoding for Single Cell Signaling Analysis and Biomarker Discovery

Published on: October 4, 2018

Area of Science:

Cell biology
Biotechnology
Cancer research

Background:

DNA barcoding advanced biomedical discovery but live imaging multiplexing is limited by fluorescent protein deconvolution.
Current microscopy techniques struggle to distinguish numerous live cells simultaneously.

Purpose of the Study:

To develop a novel method for multiplexing live imaging applications using visual barcodes.
To enable simultaneous monitoring of multiple signaling pathways at clonal resolution in mixed cell populations.

Main Methods:

Engineered visual barcodes using fluorescent proteins targeted to specific subcellular locations.
Created 'Signalome' cell-lines by mixing 12 distinct live reporter clones.
Validated barcode accuracy and robustness against cellular perturbations.

Main Results:

Demonstrated highly accurate and robust deconvolution of visual barcodes.
Successfully generated and analyzed 'Signalome' cell-lines with 12 simultaneous reporters.
Identified two distinct clusters of signaling pathways balancing growth and proliferation in cancer.

Conclusions:

Visual barcodes overcome limitations in live imaging multiplexing, enabling high-content analysis.
The 'Signalome' approach facilitates real-time, clonal-resolution monitoring of complex signaling networks.
Findings highlight growth homeostasis as a key organizing principle in cancer signaling.