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Updated: Jul 13, 2025

Genetic Barcoding with Fluorescent Proteins for Multiplexed Applications

Genetic Barcoding with Fluorescent Proteins for Multiplexed Applications

Published on: April 14, 2015

High-Content and High-Throughput Clonogenic Survival Assay Using Fluorescence Barcoding.

Haibin Qian^1,2, Selami Baglamis^2,3,4,5, Fumei Redeker^1,2

¹Department of Medical Biology, Amsterdam University Medical Centers (Location AMC), Meibergdreef 9, 1105 AZ Amsterdam, The Netherlands.

|October 14, 2023

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LeGO-CSA enhances cancer research by enabling high-content, high-throughput cell survival analysis. This new method, using fluorescently barcoded nuclei, offers greater precision than traditional Clonogenic Survival Assays (CSA).

Area of Science:

Cancer Research
Cell Biology
Drug Discovery

Background:

The Clonogenic Survival Assay (CSA) is vital for assessing cancer cell survival and proliferation.
Traditional CSA is limited by its time-consuming nature, labor intensity, and binary output.
Simultaneously achieving high-content and high-throughput analysis in cell survival assays remains a challenge.

Purpose of the Study:

To introduce LeGO-CSA, a novel method for high-content, high-throughput cell survival analysis.
To overcome the limitations of the classical CSA.
To provide a more precise and efficient tool for cancer research and drug discovery.

Main Methods:

Developed LeGO-CSA, an extension of CSA utilizing imaging of cell nuclei.
Employed fluorescent lentiviral gene ontology markers for barcoding.

Keywords:

clonogenic survival assay high-content screening high-throughput screening in 384-well plates

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Clonogenic Assay: Adherent Cells

Clonogenic Assay: Adherent Cells

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Last Updated: Jul 13, 2025

Genetic Barcoding with Fluorescent Proteins for Multiplexed Applications

Genetic Barcoding with Fluorescent Proteins for Multiplexed Applications

Published on: April 14, 2015

Competitive Genomic Screens of Barcoded Yeast Libraries

Competitive Genomic Screens of Barcoded Yeast Libraries

Published on: August 11, 2011

Clonogenic Assay: Adherent Cells

Clonogenic Assay: Adherent Cells

Published on: March 13, 2011

Validated the approach by comparing with classical CSA and conducting a small-molecule inhibitor screen.

Main Results:

LeGO-CSA enables both high-content and high-throughput analysis of cell survival and proliferation.
Classical CSA may underestimate clonogenicity.
The study revealed novel aspects of clonal cell growth.

Conclusions:

LeGO-CSA offers a robust and enhanced approach for assessing cell survival and proliferation.
The method provides increased precision and throughput compared to classical CSA.
LeGO-CSA has promising implications for accelerating drug discovery and understanding cancer biology.