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Related Experiment Video

Updated: Jun 7, 2026

Workflow for High-content, Individual Cell Quantification of Fluorescent Markers from Universal Microscope Data, Supported by Open Source Software

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Published on: December 16, 2014

A high-throughput workflow for assessing self-renewal using colony formation assays.

Majd A Al-Hamaly¹, Jessica S Blackburn²

¹Pharmacology and Nutritional Sciences, University of Kentucky, Lexington, KY 40356, USA; Markey Cancer Center, University of Kentucky, Lexington, KY 40536, USA.

Stem Cell Research

|June 27, 2025

Summary

This study presents a high-throughput colony formation assay (CFA) for T-cell acute lymphoblastic leukemia (T-ALL). The optimized workflow enables efficient drug screening by quantifying cancer cell self-renewal in a 3D matrix.

Keywords:

Colony formation assay Drug screening High-throughput Leukemia Self-renewal

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Published on: April 18, 2021

Area of Science:

Biochemistry
Cell Biology
Oncology

Background:

The colony formation assay (CFA) is crucial for evaluating cancer cell self-renewal and drug response.
Assessing self-renewal capacity is vital for understanding leukemia progression and developing effective therapies.

Purpose of the Study:

To develop and optimize a high-throughput colony formation assay (CFA) protocol for T-cell acute lymphoblastic leukemia (T-ALL).
To establish a scalable and cost-effective platform for large-scale drug screening targeting cancer cell self-renewal.

Main Methods:

A streamlined, high-throughput colony formation assay (CFA) workflow was implemented for T-ALL cells.
Colonies were cultured in a methylcellulose-based 3D matrix.
Automated image analysis was employed for robust quantification of colony number and size.

Main Results:

The protocol provides a scalable platform for assessing cancer cell self-renewal.
Automated quantification ensures reliable estimation of colony formation.
The method is cost-effective for large-scale drug screening.

Conclusions:

This optimized CFA protocol is suitable for high-throughput drug screening in T-ALL.
The assay facilitates the identification of compounds that inhibit cancer cell self-renewal.
This approach aids in prioritizing drug candidates for further in vivo validation.