Label-Free Myelin Fingerprinting: An Adult Human Post-Mortem Brain Slice Platform via Coherent Anti-Stokes Raman Spectroscopy.

Animal models and simple cell cultures often lack sufficient overlap with human physiology to study human neurological conditions effectively; one emerging alternative is the use of human organotypic slice cultures (OSCs). This study shows, using viability studies, post-mortem corpus callosum (CC) tissue OSCs cultured with a chemically defined serum-free culturing medium, maintaining viability for at least 8 days in vitro (DIV). The inclusion of CC simplifies myelin-focused research, and its potential is highlighted by visualizing molecular information within myelin by the use of label-free Coherent anti-Stokes Raman spectroscopy (CARS) imaging. This study delineates a window of 32 h after time of death (TOD) wherein myelin remains unaffected from base measurements. Finally, as a proof-of-concept, the CC tissue of a human-specific disease characterized by myelin degeneration, multiple sclerosis (MS), reveals signs of altered myelin spectral profiles when compared to non-MS myelin. In conclusion, this study provides characterization of the development of a human myelin model in combination with a CARS-focused method of analysis, establishing it as a novel toolset for future research of myelin-based neurological conditions.