Imaging Peripheral Nerve Regeneration: A New Technique for 3D Visualization of Axonal Behavior.

Leckenby, Jonathan I.; Chacon, Miranda A; Grobbelaar, Adriaan O.; Lichtman, Jeff W (2019). Imaging Peripheral Nerve Regeneration: A New Technique for 3D Visualization of Axonal Behavior. Journal of surgical research, 242, pp. 207-213. Elsevier 10.1016/j.jss.2019.04.046

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BACKGROUND Peripheral nerve assessment has traditionally been studied through histological and immunological staining techniques in a limited cross-sectional modality, making detailed analysis difficult. A new application of serial section electron microscopy is presented to overcome these limitations. METHODS Direct nerve repairs were performed on the posterior auricular nerve of transgenic YFP-H mice. Six weeks postoperatively the nerves were imaged using confocal fluorescent microscopy then excised and embedded in resin. Resin blocks were sequentially sectioned at 100 nm, and sections were serially imaged with an electron microscope. Images were aligned and autosegmented to allow for 3D reconstruction. RESULTS Basic morphometry and axonal counts were fully automated. Using full 3D reconstructions, the relationships between the axons, the Nodes of Ranvier, and Schwann cells could be fully appreciated. Interactions of individual axons with their surrounding environment could be visualized and explored in a virtual three-dimensional space. CONCLUSIONS Serial section electron microscopy allows the detailed pathway of the regenerating axon to be visualized in a 3D virtual space in comparison to isolated individual traditional histological techniques. Fully automated histo-morphometry can now give accurate axonal counts, provide information regarding the quality of nerve regeneration, and reveal the cell-to-cell interaction at a super-resolution scale. It is possible to fully visualize and "fly-through" the nerve to help understand the behavior of a regenerating axon within its environment. This technique provides future opportunities to evaluate the effect different treatment modalities have on the neuroregenerative potential and help us understand the impact different surgical techniques have when treating nerve injuries.

Item Type:

Journal Article (Original Article)

Division/Institute:

04 Faculty of Medicine > Department of Orthopaedic, Plastic and Hand Surgery (DOPH) > Clinic of Plastic and Hand Surgery
04 Faculty of Medicine > Department of Orthopaedic, Plastic and Hand Surgery (DOPH) > Clinic of Plastic and Hand Surgery > Plastic, Reconstructive and Aesthetic Surgery

UniBE Contributor:

Leckenby, Jonathan Ian and Grobbelaar, Adriaan Ockert

Subjects:

600 Technology > 610 Medicine & health

ISSN:

1095-8673

Publisher:

Elsevier

Language:

English

Submitter:

Veronika Picha

Date Deposited:

09 Jun 2020 09:51

Last Modified:

09 Jun 2020 09:59

Publisher DOI:

10.1016/j.jss.2019.04.046

PubMed ID:

31085369

Uncontrolled Keywords:

Connectomics Electron microscopy Nerve regeneration Nerve repair Peripheral nerve

BORIS DOI:

10.7892/boris.144401

URI:

https://boris.unibe.ch/id/eprint/144401

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