Mass cytometry analysis of immune cells in the brain

Area of Science:

• Neuroimmunology
• Immunology
• Cellular Biology

Background:

• Immune cells are crucial for immunological activities and interact with other cells through complex communication networks.
• The brain is an active immunological site with resident and infiltrating immune cells influencing neural functions.
• Comprehensive characterization of immune cells is essential for understanding the brain's immune network.

Purpose of the Study:

• To provide a detailed, step-by-step protocol for mass cytometry analysis of the mouse brain immune compartment.
• To enable high-dimensional single-cell analysis of brain immune cells using mass cytometry (CyTOF).
• To facilitate the study of immune cell modifications in the brain under normal and pathological conditions.

Main Methods:

• Brain perfusion, tissue extraction, and dissociation into a single-cell suspension.
• Cell staining using metal-tagged antibodies for mass cytometry (CyTOF).
• Sample reading and data analysis using SPADE and viSNE algorithms.

Main Results:

• A comprehensive mass cytometry (CyTOF) protocol for mouse brain immune cell analysis was established.
• The procedure, including staining and reading, takes less than 5 hours (excluding data analysis).
• The protocol enables high-dimensional characterization of tens of markers at the single-cell level.

Conclusions:

• Mass cytometry (CyTOF) offers a powerful approach for detailed immune profiling of the brain.
• This protocol provides a feasible method for investigating brain immune cell dynamics in various conditions.
• The technique aids in discovering novel immune populations and understanding brain immune responses.