High-content live-cell multiplex screen for chemogenomic compound annotation based on nuclear morphology
Amelie Tjaden1, Robert T Giessmann2, Stefan Knapp1
1Institute of Pharmaceutical Chemistry, Goethe University Frankfurt, Max-von-Laue-Str.9, 60438 Frankfurt, Germany; Structural Genomics Consortium, BMLS, Goethe University Frankfurt, Max-von-Laue-Str. 15, 60438 Frankfurt, Germany.
STAR Protocols
|November 1, 2022
Summary
This study presents a high-content screening method to assess compound effects on cell viability and phenotype over 48 hours. This adaptable protocol aids in drug discovery and target validation by analyzing cellular responses to small molecules.
Area of Science:
- Cell Biology
- Pharmacology
- High-Content Screening
Background:
- Well-characterized small molecules are crucial for studying cellular processes and validating drug targets.
- High-content screening (HCS) offers a powerful approach for analyzing cellular responses to chemical compounds.
- Multiplex assays allow for the simultaneous assessment of multiple cellular parameters.
Purpose of the Study:
- To describe a high-content multiplex screening protocol for evaluating cell viability over 48 hours.
- To integrate phenotypic analyses, including tubulin binding and mitochondrial content, for cellular quality control of compounds.
- To provide a scalable and adaptable live-cell based method for compound assessment.
Main Methods:
- Development of a 48-hour live-cell based high-content multiplex screen.
- Integration of cell viability assays with phenotypic feature analysis (tubulin binding, mitochondrial content).
- Utilized the CQ1 imaging system for image acquisition and CellPathfinder software for data analysis.
Main Results:
- The protocol enables simultaneous assessment of cell viability and key phenotypic features.
- The method is adaptable for various cell types and compound libraries.
- Demonstrated the utility of the screen for initial cellular quality control of diverse small molecules.
Conclusions:
- This high-content multiplex screening protocol provides a robust platform for evaluating small molecules.
- The method facilitates efficient compound characterization and target validation in a live-cell context.
- The described protocol is scalable and easily adaptable for broader applications in chemical biology and drug discovery.
