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In Vivo Two-Photon Microscopy of Single Nerve Endings in Skin
Published on: August 24, 2014
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Nerve spectroscopy: understanding peripheral nerve autofluorescence through photodynamics
Fernando Dip1,2, Rene Aleman1, Mariano Socolovsky3
1Department of General Surgery & The Bariatric and Metabolic Institute, Cleveland Clinic Florida, 2950 Cleveland. Clinic Blvd, Weston, FL, 33331, USA.
Surgical Endoscopy
|March 30, 2021
Summary
Peripheral nerve autofluorescence can visualize nerves during surgery. Nerve visualization intensity did not significantly depend on excitation wavelength or fixation methods in this ex vivo study.
Area of Science:
- Biomedical Engineering
- Surgical Technology
- Neuroscience
Background:
- Accurate identification of sensory and motor nerves is vital for preventing nerve injury during surgical procedures.
- Peripheral nerve visualization techniques are essential for improving surgical outcomes.
- Autofluorescence of nerves offers a potential method for real-time intraoperative visualization.
Purpose of the Study:
- To evaluate the feasibility of visualizing human peripheral nerves using their intrinsic autofluorescence in an ex vivo model.
- To compare the effects of three different nerve fiber fixation methods on fluorescence intensity.
- To compare the effects of three different excitation wavelength ranges on fluorescence intensity.
Main Methods:
- Human peripheral nerve samples were analyzed ex vivo.
- Nerve fibers were subjected to three fixation conditions: physiologic solution wash, formaldehyde fixation followed by wash, and formaldehyde fixation without wash.
- Nerve fibers were visualized using an inverted microscope under three different filter sets with specific excitation ranges (390-440 nm, 460-480 nm, 535-555 nm).
- Fluorescence intensity was quantified using Image-J software and analyzed with ANOVA.
Main Results:
- Fluorescence intensity ratios varied significantly between fixation groups under excitation ranges of 460-480 nm (p=0.021) and 535-555 nm (p=0.030).
- Excitation range 390-440 nm did not significantly distinguish between nerve fiber groups (p=0.39).
- All filters allowed for adequate visualization of the nerves, with Filter 1 showing higher overall intensity than Filter 3 (p<0.05).
Conclusions:
- Peripheral nerve autofluorescence intensity in an ex vivo setting is not significantly dependent on the excitation wavelength or fixation methods employed.
- These findings suggest that autofluorescence is a promising tool for nerve visualization in surgical settings.
- Further research is warranted to explore the clinical applicability of this technique for nerve-sparing surgery.
