Multiplex imaging of murine bone marrow using Phenocycler 2.0™

Area of Science:

• Biomedical research
• Cellular biology
• Tissue analysis

Background:

• Bone marrow (BM) is crucial for hematology, oncology, bone biology, and immunology research.
• Traditional methods like flow cytometry and immunofluorescence struggle to analyze unperturbed BM with multiple probes.
• Studying cellular interactions and positioning in native BM is challenging.

Purpose of the Study:

• To present a novel protocol for analyzing bone marrow (BM) cellular composition.
• To overcome limitations of traditional techniques in studying unperturbed BM.
• To enable comprehensive appreciation of diverse cell populations and their interactions within BM.

Main Methods:

• Utilizes Phenocycler 2.0™ with co-detection by indexing (CODEX) technology.
• Employs HALO 4.0™ image analysis software for data interpretation.
• Enables multiplexed analysis of cellular elements in intact BM tissue.

Main Results:

• Demonstrates the capability to study the BM in an unperturbed state.
• Facilitates detailed analysis of diverse cell populations and their spatial relationships.
• Overcomes challenges associated with traditional multi-probe fluorescent techniques.

Conclusions:

• Phenocycler 2.0™ with CODEX and HALO 4.0™ offers a powerful solution for bone marrow research.
• This protocol enhances the understanding of cellular interactions and tissue architecture.
• Enables more comprehensive studies in hematology, oncology, and immunology.