In vivo imaging of myelin in the vertebrate central nervous system using third harmonic generation microscopy

Authors

Matthew J. Farrar, Messiah UniversityFollow
Frank W. Wise, Cornell University
Joseph R. Fetcho, Cornell UniversityFollow
Chris B. Schaffer, Cornell UniversityFollow

Document Type

Article

Publication Date

3-2-2011

Abstract

Loss of myelin in the central nervous system (CNS) leads to debilitating neurological deficits. High-resolution optical imaging of myelin in the CNS of animal models is limited by a lack of in vivo myelin labeling strategies. We demonstrated that third harmonic generation (THG) microscopy - a coherent, nonlinear, dye-free imaging modality - provides micrometer resolution imaging of myelin in the mouse CNS. In fixed tissue, we found that THG signals arose from white matter tracts and were colocalized with two-photon excited fluorescence (2PEF) from a myelin-specific dye. In vivo, we used simultaneous THG and 2PEF imaging of the mouse spinal cord to resolve myelin sheaths surrounding individual fluorescently-labeled axons, and followed myelin disruption after spinal cord injury. Finally, we suggest optical mechanisms that underlie the myelin specificity of THG. These results establish THG microscopy as an ideal tool for the study of myelin loss and recovery. © 2011 by the Biophysical Society.

Comments

Farrar, M. J., Wise, F. W., Fetcho, J. R., & Schaffer, C. B. (2011). In vivo imaging of myelin in the vertebrate central nervous system using third harmonic generation microscopy. Biophysical Journal, 100(5), 1362–1371. https://doi.org/10.1016/j.bpj.2011.01.031

Recommended Citation

Farrar, Matthew J.; Wise, Frank W.; Fetcho, Joseph R.; and Schaffer, Chris B., "In vivo imaging of myelin in the vertebrate central nervous system using third harmonic generation microscopy" (2011). Educator Scholarship & Departmental Newsletters. 63.
https://mosaic.messiah.edu/mps_ed/63