Using Full-Spectrum Flow Cytometry to Phenotype Memory T and NKT Cell Subsets with Optimized Tissue-Specific...

Area of Science:

• Immunology
• Cell Biology
• Analytical Chemistry

Background:

• Full-spectrum flow cytometry is increasingly utilized for complex immune cell analysis.
• Standardized protocols are crucial for accurate panel design and ex vivo tissue analysis.
• Understanding tissue-resident immune cells requires optimized sample preparation.

Purpose of the Study:

• To provide optimized protocols for tissue processing and phenotyping of immune cells.
• To enable accurate analysis of memory T cells and natural killer T (NKT) cell subsets.
• To facilitate the study of tissue-resident memory T (T_RM) cells in various organs.

Main Methods:

• Developed optimized protocols for processing liver, lung, spleen, and lymph node tissues from mice.
• Designed a 21-color antibody panel for comprehensive immune cell phenotyping.
• Utilized full-spectrum flow cytometry for high-resolution analysis of cell subsets.

Main Results:

• Established reliable methods for isolating and analyzing memory T and NKT cell populations.
• Demonstrated the importance of controlled processing temperatures, especially for liver T_RM cells.
• Successfully identified various memory T cell subsets, including T_RM cells.

Conclusions:

• The provided protocols ensure representative ex vivo analysis of immune cells.
• Optimized tissue processing is critical for accurate phenotyping of sensitive cell subsets.
• These methods support research into adaptive immunity and tissue-resident immune cells.