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By further exploiting the qualities of the topically applied myelin dye Nile red, this technique is capable of visualizing the detailed microenvironment of peripheral nerve demyelination injury and recovery, while allowing us to obtain images of exogenous Schwann cell myelination in a living animal.<\/jats:p><\/jats:sec><jats:sec><jats:title>METHODS<\/jats:title><jats:p>The authors caused doxorubicin-induced focal demyelination in the tibial nerves of 25 Thy-1 GFP rats, of which 2 subsets (n = 10 each) received either BFP-labeled SKP-SCs or SCs to the zone of injury. Prior to acquiring images of myelin recovery in these nerves, a tibial nerve window was constructed using a silicone hemitube, a fast drying silicone polymer, and a small coverslip. This construct was then affixed to a 3D-printed nerve stage, which in turn was affixed to an external fixation\/microscope stage device. Myelin visualization was facilitated by the topical application of Nile red.<\/jats:p><\/jats:sec><jats:sec><jats:title>RESULTS<\/jats:title><jats:p>The authors reliably demonstrated intravital peripheral nerve myelin imaging with micron-level resolution and magnification, and minimal movement artifact. The detailed microenvironment of nerve remyelination can be vividly observed, while exogenously applied Schwann cells and skin-derived precursor Schwann cells can be seen myelinating axons.<\/jats:p><\/jats:sec><jats:sec><jats:title>CONCLUSIONS<\/jats:title><jats:p>Topically applied Nile red enables intravital study of myelin in the living rat PNS. Furthermore, the use of a tibial nerve window facilitates stable intravital peripheral nerve imaging, making possible high-definition spectral imaging with long acquisition times.<\/jats:p><\/jats:sec>","DOI":"10.3171\/2017.6.jns17239","type":"journal-article","created":{"date-parts":[[2018,1,19]],"date-time":"2018-01-19T14:59:33Z","timestamp":1516373973000},"page":"163-171","source":"Crossref","is-referenced-by-count":11,"title":["A novel approach to 32-channel peripheral nervous system myelin imaging in vivo, with single axon resolution"],"prefix":"10.3171","volume":"130","author":[{"given":"Joey","family":"Grochmal","sequence":"first","affiliation":[{"name":"Hotchkiss Brain Institute, Calgary;"},{"name":"Department of Clinical Neurosciences, Cumming School of Medicine, University of Calgary; and Departments of"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wulin","family":"Teo","sequence":"additional","affiliation":[{"name":"Hotchkiss Brain Institute, Calgary;"},{"name":"Neuroscience, University of Calgary, Alberta, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hardeep","family":"Gambhir","sequence":"additional","affiliation":[{"name":"Hotchkiss Brain Institute, Calgary;"},{"name":"Neuroscience, University of Calgary, Alberta, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ranjan","family":"Kumar","sequence":"additional","affiliation":[{"name":"Hotchkiss Brain Institute, Calgary;"},{"name":"Neuroscience, University of Calgary, Alberta, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jo Anne","family":"Stratton","sequence":"additional","affiliation":[{"name":"Hotchkiss Brain Institute, Calgary;"},{"name":"Neuroscience, University of Calgary, Alberta, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Raveena","family":"Dhaliwal","sequence":"additional","affiliation":[{"name":"Department of Clinical Neurosciences, Cumming School of Medicine, University of Calgary; and Departments of"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Craig","family":"Brideau","sequence":"additional","affiliation":[{"name":"Hotchkiss Brain Institute, Calgary;"},{"name":"Biomedical Engineering and"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jeff","family":"Biernaskie","sequence":"additional","affiliation":[{"name":"Hotchkiss Brain Institute, Calgary;"},{"name":"Neuroscience, University of Calgary, Alberta, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Peter K.","family":"Stys","sequence":"additional","affiliation":[{"name":"Hotchkiss Brain Institute, Calgary;"},{"name":"Department of Clinical Neurosciences, Cumming School of Medicine, University of Calgary; and Departments of"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rajiv","family":"Midha","sequence":"additional","affiliation":[{"name":"Hotchkiss Brain Institute, Calgary;"},{"name":"Department of Clinical Neurosciences, Cumming School of Medicine, University of Calgary; and Departments of"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1754","reference":[{"key":"b1","series-title":"Proc Natl Acad Sci U S A","doi-asserted-by":"publisher","first-page":"6282","DOI":"10.1073\/pnas.1015239108","article-title":"In vivo imaging reveals a phase-specific role of STAT3 during central and peripheral nervous system axon regeneration","volume":"108","author":"Bareyre FM","year":"2011","unstructured":"Bareyre FM, Garzorz N, Lang C, Misgeld T, B\u00fcning H, Kerschensteiner M: In vivo imaging reveals a phase-specific role of STAT3 during central and peripheral nervous system axon regeneration. 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