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Updated: Jun 4, 2026
Bioluminescence and Near-infrared Imaging of Optic Neuritis and Brain Inflammation in the EAE Model of Multiple Sclerosis in Mice
Published on: March 1, 2017
Multimodal coherent anti-Stokes Raman scattering microscopy reveals microglia-associated myelin and axonal
Jaime Imitola1, Daniel Côté, Stine Rasmussen
1Brigham and Women's Hospital, Center for Neurologic Diseases, Partner Multiple Sclerosis Center, Harvard Medical School, Department of Neurology, Boston, Massachusetts 02115, USA.
Abstract:
Myelin loss and axonal degeneration predominate in many neurological disorders; however, methods to visualize them simultaneously in live tissue are unavailable. We describe a new imaging strategy combining video rate reflectance and fluorescence confocal imaging with coherent anti-Stokes Raman scattering (CARS) microscopy tuned to CH(2) vibration of myelin lipids, applied in live tissue of animals with chronic experimental autoimmune encephalomyelitis (EAE). Our method allows monitoring over time of demyelination and neurodegeneration in brain slices with high spatial resolution and signal-to-noise ratio. Local areas of severe loss of lipid signal indicative of demyelination and loss of the reflectance signal from axons were seen in the corpus callosum and spinal cord of EAE animals. Even in myelinated areas of EAE mice, the intensity of myelin lipid signals is significantly reduced. Using heterozygous knock-in mice in which green fluorescent protein replaces the CX(3)CR1 coding sequence that labels central nervous system microglia, we find areas of activated microglia colocalized with areas of altered reflectance and CARS signals reflecting axonal injury and demyelination. Our data demonstrate the use of multimodal CARS microscopy for characterization of demyelinating and neurodegenerative pathology in a mouse model of multiple sclerosis, and further confirm the critical role of microglia in chronic inflammatory neurodegeneration.
Insights
This study introduces a novel imaging technique to simultaneously visualize myelin and axonal damage in live neurological tissue. This method aids in understanding demyelinating diseases like multiple sclerosis and the role of microglia.
Area of Science:
- Neuroscience
- Biomedical Imaging
- Pathology
Background:
- Neurological disorders often involve myelin loss and axonal degeneration.
- Simultaneous visualization of these pathologies in live tissue remains a challenge.
Purpose of the Study:
- To develop and validate a new imaging strategy for real-time monitoring of demyelination and neurodegeneration.
- To investigate the role of microglia in chronic inflammatory neurodegeneration.
Main Methods:
- Combined video rate reflectance, fluorescence confocal imaging, and coherent anti-Stokes Raman scattering (CARS) microscopy.
- Applied to live brain slices from mice with experimental autoimmune encephalomyelitis (EAE).
- CARS microscopy tuned to CH(2) vibration of myelin lipids.
Main Results:
- Successfully visualized demyelination and axonal injury in live EAE mouse tissue with high resolution.
- Observed reduced myelin lipid signals and axonal reflectance in affected areas.
- Activated microglia colocalized with regions of demyelination and axonal injury.
Conclusions:
- Multimodal CARS microscopy is effective for characterizing demyelinating and neurodegenerative pathology in EAE models.
- Highlights the significant role of microglia in chronic inflammatory neurodegeneration.
- Provides a new tool for studying live tissue in neurological disease research.
