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High-Content Screening Assay for the Identification of Antibody-Dependent Cellular Cytotoxicity Modifying Compounds
Published on: August 18, 2023
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High Content, Phenotypic Assays and Screens for Compounds Modulating Cellular Processes in Primary Neurons
Miklos Kepiro1, Boglarka H Varkuti1, Ronald L Davis1
1Department of Neuroscience, The Scripps Research Institute Florida, Jupiter, FL, United States.
Methods in Enzymology
|November 5, 2018
Summary
High-content phenotypic screening uses automated microscopy and image analysis to study cellular systems biology. This approach, applied to mitochondrial dynamics in neurons, enables high-throughput identification of cellular perturbations.
Area of Science:
- Cellular biology
- Systems biology
- High-content screening
Background:
- High-content screening enables cellular systems biology by analyzing images of labeled cells.
- Automated microscopy and image processing are key components of these assays.
Purpose of the Study:
- To describe the advantages and limitations of high-content assays.
- To detail methods for assaying mitochondrial dynamics in primary neurons.
- To provide an open-source CellProfiler pipeline for image analysis.
Main Methods:
- Utilizing cells with fluorescently labeled molecules or organelles in microplates.
- Employing automated confocal microscopy for image acquisition.
- Applying image processing, feature extraction, and data analysis pipelines.
Main Results:
- Demonstrated the application of high-content screening to assay mitochondrial dynamics.
- Provided detailed culturing, imaging, and data analysis techniques.
- Developed an open-source CellProfiler pipeline for high-throughput image analysis.
Conclusions:
- High-content screening is a powerful tool for systems biology at the cellular level.
- The described methods and open-source pipeline facilitate the study of mitochondrial dynamics.
- This approach allows for the identification of cellular perturbations with high throughput.
Keywords:
Automated image analysisCellProfilerHigh content phenotypic screenHit selectionMitochondrial dynamicsMitochondrial morphologyPintoolPrimary neuronsMore Related Videos
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