Spatial immunophenotyping of FFPE tissues by imaging mass cytometry

Area of Science:

• Immunology
• Biotechnology
• Pathology

Background:

• Tissue immune microenvironments are dynamic and crucial for response outcomes.
• Immune cell localization, not just composition, dictates response.
• Conventional methods like immunohistochemistry (IHC) have limitations in multiplexing.

Purpose of the Study:

• To present a protocol for immunophenotyping formalin-fixed, paraffin-embedded (FFPE) tissues using imaging mass cytometry (IMC).
• To overcome the target number limitations of traditional imaging techniques.
• To facilitate the characterization of immune niches and their alterations.

Main Methods:

• Developed a protocol for IMC-based immunophenotyping of FFPE tissues.
• Included detailed staining procedures and an example murine antibody panel.
• Provided suggestions for initial image analysis.

Main Results:

• IMC allows simultaneous detection of up to 40 targets with 1-μm resolution.
• The protocol is applicable to FFPE tissues, including clinically relevant samples.
• Demonstrated a workflow for characterizing immune niches.

Conclusions:

• The presented IMC protocol offers a powerful method for detailed immune cell analysis in FFPE tissues.
• This approach enhances the study of immune responses, disease progression, and therapeutic interventions.
• Facilitates deeper understanding of immune cell interactions within tissue microenvironments.