Nerve spectroscopy: understanding peripheral nerve autofluorescence through photodynamics.


Journal

Surgical endoscopy
ISSN: 1432-2218
Titre abrégé: Surg Endosc
Pays: Germany
ID NLM: 8806653

Informations de publication

Date de publication:
12 2021
Historique:
received: 30 07 2020
accepted: 03 12 2020
pubmed: 31 3 2021
medline: 6 1 2022
entrez: 30 3 2021
Statut: ppublish

Résumé

Being able to accurately identify sensory and motor nerves is crucial during surgical procedures to prevent nerve injury. We aimed to (1) evaluate the feasibility of performing peripheral human nerve visualization utilizing nerves' own autofluorescence in an ex-vivo model; (2) compare the effect of three different nerve fiber fixation methods on the intensity of fluorescence, indicated as the intensity ratio; and (3) similarly compare three different excitation ranges. Samples from various human peripheral nerves were selected postoperatively. Nerve fibers were divided into three groups: Group A nerve fibers were washed with a physiologic solution; Group B nerve fibers were fixated with formaldehyde for 6 h first, and then washed with a physiologic solution; Group C nerve fibers were fixated with formaldehyde for six hours, but not washed afterwards. An Olympus IX83 inverted microscope was used for close-up image evaluation. Nerve fibers were exposed to white-light wavelength spectrums for a specific time frame prior to visualization under three different filters-Filter 1-LF405-B-OMF Semrock; Filter 2-U-MGFP; Filter 3-U-MRFPHQ Olympus, with excitation ranges of 390-440, 460-480, and 535-555, respectively. The fluorescence intensity of all images was subsequently analyzed using Image-J Software, and results compared by analysis of variance (ANOVA). The intensity ratios observed with Filter 1 failed to distinguish the different nerve fiber groups (p = 0.39). Conversely, the intensity ratios seen under Filters 2 and 3 varied significantly between the three nerve-fiber groups (p = 0.021, p = 0.030, respectively). The overall intensity of measurements was greater with Filter 1 than Filter 3 (p < 0.05); however, all nerves were well visualized by all filters. The current results on ex vivo peripheral nerve fiber autofluorescence suggest that peripheral nerve fiber autofluorescence intensity does not greatly depend upon the excitation wavelength or fixation methods used in an ex vivo setting. Implications for future nerve-sparing surgery are discussed.

Sections du résumé

BACKGROUND
Being able to accurately identify sensory and motor nerves is crucial during surgical procedures to prevent nerve injury. We aimed to (1) evaluate the feasibility of performing peripheral human nerve visualization utilizing nerves' own autofluorescence in an ex-vivo model; (2) compare the effect of three different nerve fiber fixation methods on the intensity of fluorescence, indicated as the intensity ratio; and (3) similarly compare three different excitation ranges.
METHODS
Samples from various human peripheral nerves were selected postoperatively. Nerve fibers were divided into three groups: Group A nerve fibers were washed with a physiologic solution; Group B nerve fibers were fixated with formaldehyde for 6 h first, and then washed with a physiologic solution; Group C nerve fibers were fixated with formaldehyde for six hours, but not washed afterwards. An Olympus IX83 inverted microscope was used for close-up image evaluation. Nerve fibers were exposed to white-light wavelength spectrums for a specific time frame prior to visualization under three different filters-Filter 1-LF405-B-OMF Semrock; Filter 2-U-MGFP; Filter 3-U-MRFPHQ Olympus, with excitation ranges of 390-440, 460-480, and 535-555, respectively. The fluorescence intensity of all images was subsequently analyzed using Image-J Software, and results compared by analysis of variance (ANOVA).
RESULTS
The intensity ratios observed with Filter 1 failed to distinguish the different nerve fiber groups (p = 0.39). Conversely, the intensity ratios seen under Filters 2 and 3 varied significantly between the three nerve-fiber groups (p = 0.021, p = 0.030, respectively). The overall intensity of measurements was greater with Filter 1 than Filter 3 (p < 0.05); however, all nerves were well visualized by all filters.
CONCLUSION
The current results on ex vivo peripheral nerve fiber autofluorescence suggest that peripheral nerve fiber autofluorescence intensity does not greatly depend upon the excitation wavelength or fixation methods used in an ex vivo setting. Implications for future nerve-sparing surgery are discussed.

Identifiants

pubmed: 33782757
doi: 10.1007/s00464-020-08227-7
pii: 10.1007/s00464-020-08227-7
doi:

Types de publication

Journal Article

Langues

eng

Sous-ensembles de citation

IM

Pagination

7104-7111

Informations de copyright

© 2021. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.

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Auteurs

Fernando Dip (F)

Department of General Surgery & The Bariatric and Metabolic Institute, Cleveland Clinic Florida, 2950 Cleveland. Clinic Blvd, Weston, FL, 33331, USA.
Department of General Surgery, Hospital de Clínicas José de San Martín, University of Buenos Aires, Av. Córdoba 2351, C1121ABJ, CABA, Argentina.

Rene Aleman (R)

Department of General Surgery & The Bariatric and Metabolic Institute, Cleveland Clinic Florida, 2950 Cleveland. Clinic Blvd, Weston, FL, 33331, USA.

Mariano Socolovsky (M)

Department of Neurosurgery, Hospital de Clínicas José de San Martín, University of Buenos Aires, Buenos Aires, Argentina.

Nerina Villalba (N)

IBCN, Facultad de Medicina, University of Buenos Aires, Buenos Aires, Argentina.

Jorge Falco (J)

Department of General Surgery, Hospital de Clínicas José de San Martín, University of Buenos Aires, Av. Córdoba 2351, C1121ABJ, CABA, Argentina.

Emanuele Lo Menzo (EL)

Department of General Surgery & The Bariatric and Metabolic Institute, Cleveland Clinic Florida, 2950 Cleveland. Clinic Blvd, Weston, FL, 33331, USA.

Kevin P White (KP)

Science Right Research Consulting Inc, 195 Dufferin Ave., Suite 605, London, ON, N6A 1K7, Canada.

Raul J Rosenthal (RJ)

Department of General Surgery & The Bariatric and Metabolic Institute, Cleveland Clinic Florida, 2950 Cleveland. Clinic Blvd, Weston, FL, 33331, USA. ROSENTR@ccf.org.

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