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

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.

journal_name

Biophys J

journal_title

Biophysical journal

authors

Farrar MJ,Wise FW,Fetcho JR,Schaffer CB

doi

10.1016/j.bpj.2011.01.031

subject

Has Abstract

pub_date

2011-03-02 00:00:00

pages

1362-71

issue

5

eissn

0006-3495

issn

1542-0086

pii

S0006-3495(11)00114-7

journal_volume

100

pub_type

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