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Using High Content Imaging to Quantify Target Engagement in Adherent Cells
Published on: November 29, 2018
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Cell Proliferation High-Content Screening on Adherent Cell Cultures
Pau Carrillo-Barberà1,2,3, Jose M Morante-Redolat1,2,3, José F Pertusa4
1Departamento de Biología Celular, Biología Funcional y Antropología Física, Universitat de València, Burjassot, Spain.
Methods in Molecular Biology (Clifton, N.J.)
|August 22, 2019
Summary
This study presents a high-content proliferation assay for adherent cells using nucleoside analogues like EdU. The method quantifies cell cycle phases (G0-M) and proliferation rates in specific cell subpopulations.
Area of Science:
- Cell Biology
- Molecular Biology
- Biotechnology
Background:
- Cell proliferation is crucial for development and disease.
- Accurate measurement of cell cycle progression is essential for biological research.
- Existing methods may lack the throughput or specificity required for complex analyses.
Purpose of the Study:
- To describe a high-content proliferation assay pipeline for adherent cell cultures.
- To enable detailed analysis of cell cycle phases and proliferation rates.
- To facilitate the identification of specific cell subpopulations based on nuclear markers.
Main Methods:
- Utilized pulse-chase experiments with nucleoside analogues (BrdU/EdU) to label cells in S phase.
- Developed a high-throughput imaging and non-supervised ImageJ macroinstruction for data analysis.
- Integrated nuclear segmentation, nucleoside analogue detection, and additional nuclear marker signal measurement.
Main Results:
- Successfully quantified the percentage of cells in different cell cycle phases (G0, G1, S, G2, M).
- Enabled estimation of proliferation (S phase) rates within specific cell subpopulations.
- Demonstrated the pipeline's capability for high-content analysis of cell proliferation.
Conclusions:
- The described pipeline offers a robust method for high-content proliferation analysis in adherent cells.
- This assay allows for precise determination of cell cycle distribution and proliferation rates.
- The method is adaptable for studying proliferation in distinct cell subpopulations identified by nuclear markers.
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