Abstract
Nerve-sparing surgery is increasingly being applied to avoid functional deficits of the limbs and organs following surgery. Peripheral nerves that should be preserved are, however, sometimes misidentified due to similarity of shape and color to non-nerve tissues. To avoid misidentification of peripheral nerves, development of an in situ nerve detection method is desired. In this study, we report the label-free detection of ex vivo peripheral nerves of Wistar rats by using Raman spectroscopy. We obtained Raman spectra of peripheral nerves (myelinated and unmyelinated nerves) and their adjacent tissues of Wistar rats without any treatment such as fixation and/or staining. For the identification of tissue species and further analysis of spectral features, we proposed a principal component regression-based discriminant analysis with representative Raman spectra of peripheral nerves and their adjacent tissues. Our prediction model selectively detected myelinated nerves and unmyelinated nerves of Wistar rats with respective sensitivities of 95.5% and 88.3% and specificities of 99.4% and 93.5%. Furthermore, important spectral features for the identification of tissue species were revealed by detailed analysis of principal components of representative Raman spectra of tissues. Our proposed approach may provide a unique and powerful tool for peripheral nerve detection for nerve-sparing surgery in the future.
Highlights
We have provided a proof-of-principle demonstration of selective detection of peripheral nerves including myelinated and unmyelinated nerves against adjacent tissues by using dried microsection samples after a frozen-thawed procedure[23]
We investigated the feasibility of peripheral nerve detection with Raman spectroscopy by using fresh ex vivo macrosamples from Wistar rats
A principal component regression (PCR)-based discriminant analysis was performed for the selective detection and further spectral analysis of peripheral nerves against adjacent tissues, and the detection power of peripheral nerves was examined
Summary
We have provided a proof-of-principle demonstration of selective detection of peripheral nerves including myelinated and unmyelinated nerves against adjacent tissues by using dried microsection samples after a frozen-thawed procedure[23]. In such dried microsection samples, cross-sections of nerve fibers were directly examined by means of Raman spectroscopy. We investigated the feasibility of peripheral nerve detection with Raman spectroscopy by using fresh ex vivo macrosamples from Wistar rats. We examined myelinated nerve, unmyelinated nerve, adipose tissue, collagenous tissue, and skeletal muscle without any treatments such as fixation, staining, or microsectioning. A principal component regression (PCR)-based discriminant analysis was performed for the selective detection and further spectral analysis of peripheral nerves against adjacent tissues, and the detection power of peripheral nerves was examined
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