Protocol for multispectral imaging on cryosections to map myeloid cell heterogeneity in its spatial context

Area of Science:

• Immunology
• Cell Biology
• Biotechnology

Background:

• Advanced techniques like single-cell RNA sequencing and mass cytometry offer high-resolution cell identification.
• These methods often sacrifice crucial cellular and molecular context within tissues.
• Understanding cell heterogeneity in its native environment is vital for disease research.

Purpose of the Study:

• To present a detailed protocol for in situ multispectral imaging.
• To enable the mapping of myeloid cell heterogeneity directly within tissue cryosections.
• To provide a method that retains cellular and molecular context.

Main Methods:

• Protocol development for sequential tissue cryosectioning.
• Antibody titration and spectral library construction.
• Detailed procedures for multispectral imaging and data preparation for analysis.

Main Results:

• Established a comprehensive protocol for in situ multispectral imaging of tissue cryosections.
• Demonstrated the ability to map myeloid cell heterogeneity while preserving tissue context.
• Provided a framework for downstream analysis of complex cellular landscapes.

Conclusions:

• In situ multispectral imaging is a powerful approach to study cell heterogeneity in its native context.
• This protocol facilitates detailed analysis of myeloid cell populations in various tissue types.
• The method complements existing single-cell technologies by preserving spatial information.