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Research Article
Iván Coto Hernández
Massachusetts Eye and Ear Infirmary
Corresponding Author
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Traditional histopathologic evaluation of peripheral nerve employs brightfield microscopy with diffraction limited resolution of ~ 250 nm. Though electron microscopy yields nanoscale resolution of the nervous system, it is resource-intensive and incompatible with life. Super-resolution microscopy (SRM) comprises a set of imaging techniques permitting unprecedented resolution of fluorescent objects using visible light. The advent of SRM has transformed biomedical science in establishing non-toxic means for investigation of nanoscale cellular structures. Herein, sciatic nerve sections from GFP-variant expressing mice, and regenerating human nerve from cross-facial autografts labelled with a myelin-specific fluorescent dye were imaged by super-resolution radial fluctuation microscopy, stimulated emission depletion microscopy, and structured illumination microscopy. Super-resolution imaging of axial cryosections of murine sciatic nerves demonstrated robust visualization of myelinated and unmyelinated axons. Super-resolution imaging of axial cryosections of human cross-facial nerve grafts demonstrated enhanced resolution of small-calibre thinly-myelinated regenerating motor axons. The utility of SRM in imaging of mammalian cranial and peripheral nerves is demonstrated. The increase in contrast and structural clarity achievable with super-resolution techniques enables visualization of unmyelinated axons, regenerating axons, cytoskeleton ultrastructure, and neuronal appendages using light microscopes.
Keywords
Super-Resolution Microscopy, Microscopy, Fluorescence, Nerve Regeneration, Peripheral Nervous System, Histology, Facial Paralysis
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No competing interests reported.
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CURRENT STATUS: Under Revision
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Posted 25 Jan, 2021
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Editorial decision: Major revision
On 21 Apr, 2021
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Received 12 Apr, 2021
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On 29 Mar, 2021
Reviews received
Received 08 Mar, 2021
Reviewers agreed
On 25 Feb, 2021
Reviewers invited
Invitations sent on 21 Feb, 2021
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On 15 Feb, 2021
Editor invited
On 27 Jan, 2021
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On 21 Jan, 2021
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Subject Areas
Neurology
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This preprint is under consideration at Scientific Reports. A preprint is a preliminary version of a manuscript that has not completed peer review at a journal. Research Square does not conduct peer review prior to posting preprints. The posting of a preprint on this server should not be interpreted as an endorsement of its validity or suitability for dissemination as established information or for guiding clinical practice.
Learn more about In Review
Research Article
Iván Coto Hernández
Massachusetts Eye and Ear Infirmary
Corresponding Author
Badges
Prescreen
This manuscript passed our Prescreen assessment
Complete author information
Appropriate declaration statements
Potential risks to human health
Prescreen
Peer Review Timeline
CURRENT STATUS: Under Revision
Version 1
Posted 25 Jan, 2021
No community comments so far
Editorial decision: Major revision
On 21 Apr, 2021
Reviews received
Received 12 Apr, 2021
Reviewers agreed
On 29 Mar, 2021
Reviews received
Received 08 Mar, 2021
Reviewers agreed
On 25 Feb, 2021
Reviewers invited
Invitations sent on 21 Feb, 2021
Editor assigned
On 15 Feb, 2021
Editor invited
On 27 Jan, 2021
Submission checks complete
On 21 Jan, 2021
First submitted
On 05 Jan, 2021
Metrics
Comments: 0
PDF Downloads: ...
HTML Views: ...
Subject Areas
Neurology
License:
This work is licensed under a CC BY 4.0 License. Read Full License
Traditional histopathologic evaluation of peripheral nerve employs brightfield microscopy with diffraction limited resolution of ~ 250 nm. Though electron microscopy yields nanoscale resolution of the nervous system, it is resource-intensive and incompatible with life. Super-resolution microscopy (SRM) comprises a set of imaging techniques permitting unprecedented resolution of fluorescent objects using visible light. The advent of SRM has transformed biomedical science in establishing non-toxic means for investigation of nanoscale cellular structures. Herein, sciatic nerve sections from GFP-variant expressing mice, and regenerating human nerve from cross-facial autografts labelled with a myelin-specific fluorescent dye were imaged by super-resolution radial fluctuation microscopy, stimulated emission depletion microscopy, and structured illumination microscopy. Super-resolution imaging of axial cryosections of murine sciatic nerves demonstrated robust visualization of myelinated and unmyelinated axons. Super-resolution imaging of axial cryosections of human cross-facial nerve grafts demonstrated enhanced resolution of small-calibre thinly-myelinated regenerating motor axons. The utility of SRM in imaging of mammalian cranial and peripheral nerves is demonstrated. The increase in contrast and structural clarity achievable with super-resolution techniques enables visualization of unmyelinated axons, regenerating axons, cytoskeleton ultrastructure, and neuronal appendages using light microscopes.
Figure 1
Figure 2
Figure 3
Figure 4
No competing interests reported.
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