Multimodal coherent anti-Stokes Raman scattering microscopy reveals microglia-associated myelin and axonal dysfunction in multiple sclerosis-like lesions in mice

Jaime Imitola; Stine Rasmussen; Yingru Liu; Tanuja Chitnis; Samia Khoury; Daniel Côté; X. Sunney Xie; Charles P. Lin; Richard L. Sidman

doi:10.1117/1.3533312

1 February 2011 Multimodal coherent anti-Stokes Raman scattering microscopy reveals microglia-associated myelin and axonal dysfunction in multiple sclerosis-like lesions in mice

Jaime Imitola, Stine Rasmussen, Yingru Liu, Tanuja Chitnis, Samia Khoury, Daniel Côté, X. Sunney Xie, Charles P. Lin, Richard L. Sidman

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Journal of Biomedical Optics, Vol. 16, Issue 2, 021109 (February 2011). https://doi.org/10.1117/1.3533312

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 CH₂vibration 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 CX₃CR1 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.

©(2011) Society of Photo-Optical Instrumentation Engineers (SPIE)

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Jaime Imitola, Stine Rasmussen, Yingru Liu, Tanuja Chitnis, Samia Khoury, Daniel Côté, X. Sunney Xie, Charles P. Lin, and Richard L. Sidman "Multimodal coherent anti-Stokes Raman scattering microscopy reveals microglia-associated myelin and axonal dysfunction in multiple sclerosis-like lesions in mice," Journal of Biomedical Optics 16(2), 021109 (1 February 2011). https://doi.org/10.1117/1.3533312

Published: 1 February 2011

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