High Dimensional Proteomic Multiplex Imaging of the Central Nervous System Using the COMET^™ System

Area of Science:

• Neuroscience
• Biotechnology
• Medical Imaging

Background:

• Sequential multiplex methods require antibody modification, limiting reagent use and necessitating validation.
• Fixed antibody panels face limitations like spectral overlap, steric hindrance, and tissue degradation.
• Current methods for CNS cell profiling are often low-throughput and lack flexibility.

Purpose of the Study:

• To develop and validate a CNS-specific antibody library and analysis platform for high-throughput cell profiling.
• To leverage the automated Lunaphore COMET™ platform for label-free primary antibodies and secondary antibody detection.
• To address the unmet need for flexible and efficient spatial profiling in the human central nervous system.

Main Methods:

• Utilized the automated Lunaphore COMET™ platform for staining and imaging formalin-fixed paraffin-embedded (FFPE) CNS histology slides.
• Developed and validated a CNS-specific antibody library, including strategies for managing CNS autofluorescence.
• Employed off-the-shelf label-free primary antibodies and fluorophore-labeled secondary antibodies for automated slide processing.

Main Results:

• Successfully cataloged and validated a large series of antibodies for CNS spatial profiling on the COMET™ platform.
• Achieved subcellular-level visualization and quantification of cell populations and networks within the CNS.
• Demonstrated the capability to analyze up to 40 analytes per specimen, with significant clinical utility for limited CNS biopsy samples.

Conclusions:

• Established a general framework for imaging and quantifying cell populations and networks within the CNS.
• Provided the scientific community with optimized standard operating procedures for CNS spatial profiling.
• The developed platform offers a flexible and efficient solution for advancing neuroscience research and diagnostics.