A novel approach to 32-channel peripheral nervous system myelin imaging in vivo, with single axon resolution
Autor: | Hardeep Gambhir, Peter K. Stys, Ranjan Kumar, Joey Grochmal, Wulin Teo, Rajiv Midha, Jo Anne Stratton, Craig Brideau, Jeff Biernaskie, Raveena Dhaliwal |
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Rok vydání: | 2018 |
Předmět: |
Male
0301 basic medicine Intravital Microscopy Confocal Schwann cell Rats Sprague-Dawley 03 medical and health sciences chemistry.chemical_compound Myelin 0302 clinical medicine Peripheral Nerve Injuries Oxazines medicine Animals Remyelination Axon Tibial nerve Myelin Sheath Fluorescent Dyes business.industry Nile red General Medicine Axons Rats Disease Models Animal 030104 developmental biology medicine.anatomical_structure chemistry Peripheral nervous system Schwann Cells Tibial Nerve business 030217 neurology & neurosurgery Biomedical engineering |
Zdroj: | Journal of Neurosurgery. 130:163-171 |
ISSN: | 1933-0693 0022-3085 |
Popis: | OBJECTIVEIntravital spectral imaging of the large, deeply situated nerves in the rat peripheral nervous system (PNS) has not been well described. Here, the authors have developed a highly stable platform for performing imaging of the tibial nerve in live rodents, thus allowing the capture of high-resolution, high-magnification spectral images requiring long acquisition times. 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.METHODSThe 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.RESULTSThe 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.CONCLUSIONSTopically 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. |
Databáze: | OpenAIRE |
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