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Mass cytometry as a tool in target validation and drug discovery
Edmund C R Watson1, Warren Baker1, David Ahern2
1Botnar Research Centre, NIHR Biomedical Research Centre, Nuffield Department of Orthopedics, Rheumatology and Musculoskeletal Sciences, University of Oxford, United Kingdom; Oxford Translational Myeloma Centre, Botnar Research Centre, University of Oxford, United Kingdom.
Methods in Enzymology
|October 20, 2023
Summary
Mass cytometry offers detailed single-cell analysis for drug impact studies. This chapter details experimental planning and execution, showcasing its utility in diverse biological investigations.
Area of Science:
- Biotechnology
- Cell Biology
- Immunology
Background:
- Mass cytometry enables high-dimensional, single-cell analysis by combining mass spectrometry with flow cytometry.
- It is valuable for understanding heterogeneous drug effects across different cell subtypes.
- This chapter provides a comprehensive guide to mass cytometry experimental design and execution.
Approach:
- Detailed review of mass cytometry platform technical aspects and limitations.
- Step-by-step methodology modules covering staining, data acquisition, and cleaning.
- Three distinct experimental examples demonstrating core mass cytometry applications.
Key Points:
- Experiment 1: Validation and titration of metal-conjugated antibody reporters.
- Experiment 2: Assessing kinase inhibitor effects on myeloma cell cycle and apoptosis.
- Experiment 3: Analyzing drug-induced transcription factor changes across cellular compartments in PBMCs.
Conclusions:
- Mass cytometry is a powerful tool for dissecting complex cellular responses to stimuli.
- The described methodologies facilitate robust experimental design and data interpretation.
- This approach enhances understanding of drug mechanisms and cellular heterogeneity.
