Histochemistry and Cell Biology

, Volume 139, Issue 1, pp 181–193 | Cite as

Label-free detection of peripheral nerve tissues against adjacent tissues by spontaneous Raman microspectroscopy

  • Takeo Minamikawa
  • Yoshinori Harada
  • Noriaki Koizumi
  • Koji Okihara
  • Kazumi Kamoi
  • Akio Yanagisawa
  • Tetsuro Takamatsu
Original Paper

Abstract

Detection of peripheral nerve tissues during surgery is required to avoid neural disturbance following surgery as an aspect of realizing better functional outcome. We provide a proof-of-principle demonstration of a label-free detection technique of peripheral nerve tissues, including myelinated and unmyelinated nerves, against adjacent tissues that employ spontaneous Raman microspectroscopy. To investigate the Raman spectral features of peripheral nerves in detail, we used unfixed sectioned samples. Raman spectra of myelinated nerve, unmyelinated nerve, fibrous connective tissue, skeletal muscle, tunica media of blood vessel, and adipose tissue of Wistar rats were analyzed, and Raman images of the tissue distribution were constructed using the map of the ordinary least squares regression (OLSR) estimates. We found that nerve tissues exhibited a specific Raman spectrum arising from axon or myelin sheath, and that the nerve tissues can be selectively detected against the other tissues. Moreover, myelinated and unmyelinated nerves can be distinguished by the intensity differences of 2,855 cm−1, and 2,945 cm−1, which are mainly derived from lipid and protein contents of nerve fibers. We applied this method to unfixed section samples of human periprostatic tissues excised from prostatic cancer patients. Myelinated nerves, unmyelinated nerves, fibrous connective tissues, and adipose tissues of the periprostatic tissues were separately detected by OLSR analysis. These results suggest the potential of the Raman spectroscopic observation for noninvasive and label-free nerve detection, and we expect this method could be a key technique for nerve-sparing surgery.

Keywords

Raman microspectroscopy Unmyelinated and myelinated nerves Tissue imaging Nerve detection Nerve-sparing surgery 

Notes

Acknowledgments

We thank T. Okuda and T. Kawamura of the Department of Pathology and Cell Regulation, Kyoto Prefectural University of Medicine, for histological staining. A portion of this work was supported by a Grant-in-Aid for Scientific Research from the Ministry of Education, Culture, Sports, Science and Technology (MEXT), and Research for Promoting Technological Seeds from Japan Science and Technology Agency (JST). One of the authors (T.M.) acknowledges support by a Grant-in-Aid for JSPS Fellows from the Japan Society for the Promotion of Science (JSPS).

Supplementary material

418_2012_1015_MOESM1_ESM.pdf (70 kb)
Supplementary material 1 (PDF 69 kb)

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Copyright information

© Springer-Verlag 2012

Authors and Affiliations

  • Takeo Minamikawa
    • 1
  • Yoshinori Harada
    • 1
  • Noriaki Koizumi
    • 1
    • 2
  • Koji Okihara
    • 3
  • Kazumi Kamoi
    • 3
  • Akio Yanagisawa
    • 4
  • Tetsuro Takamatsu
    • 1
  1. 1.Department of Pathology and Cell RegulationGraduate School of Medical Science, Kyoto Prefectural University of MedicineKyotoJapan
  2. 2.Division of Digestive Surgery, Department of SurgeryGraduate School of Medical Science, Kyoto Prefectural University of MedicineKyotoJapan
  3. 3.Department of UrologyGraduate School of Medical Science, Kyoto Prefectural University of MedicineKyotoJapan
  4. 4.Department of Surgical PathologyGraduate School of Medical Science, Kyoto Prefectural University of MedicineKyotoJapan

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