Label-free detection of peripheral nerve tissues against adjacent tissues by spontaneous Raman microspectroscopy
- 764 Downloads
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.
KeywordsRaman microspectroscopy Unmyelinated and myelinated nerves Tissue imaging Nerve detection Nerve-sparing surgery
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).
- Gao L, Zhou H, Thrall MJ, Li F, Yang Y, Wang Z, Luo P, Wong KK, Palapattu GS, Wong ST (2011) Label-free high-resolution imaging of prostate glands and cavernous nerves using coherent anti-Stokes Raman scattering microscopy. Biomedical Opt Express 2(4):915–926. doi: 10.1364/BOE.2.000915 CrossRefGoogle Scholar
- Han M, Kim C, Mozer P, Schafer F, Badaan S, Vigaru B, Tseng K, Petrisor D, Trock B, Stoianovici D (2011) Tandem-robot assisted laparoscopic radical prostatectomy to improve the neurovascular bundle visualization: a feasibility study. Urology 77(2):502–506. doi: 10.1016/j.urology.2010.06.064 PubMedCrossRefGoogle Scholar
- Harada Y, Takamatsu T (2012) Raman molecular imaging of cells and tissues: towards functional diagnostic imaging without labeling. Curr Pharm Biotechnol (E-pub ahead of print)Google Scholar
- Hashimoto K, Hisasue S, Masumori N, Kobayashi K, Kato R, Fukuta F, Takahashi A, Hasegawa T, Tsukamoto T (2010) Clinical safety and feasibility of a newly developed, simple algorithm for decision-making on neurovascular bundle preservation in radical prostatectomy. Jpn J Clin Oncol 40(4):343–348. doi: 10.1093/jjco/hyp157 PubMedCrossRefGoogle Scholar
- Montorsi F, Salonia A, Suardi N, Gallina A, Zanni G, Briganti A, Deho F, Naspro R, Farina E, Rigatti P (2005) Improving the preservation of the urethral sphincter and neurovascular bundles during open radical retropubic prostatectomy. Eur Urol 48(6):938–945. doi: 10.1016/j.eururo.2005.09.004 PubMedCrossRefGoogle Scholar
- Nakano K, Harada Y, Yamaoka Y, Miyawaki K, Imaizumi K, Takaoka H, Nakaoka M, Wakabayashi N, Yoshikawa T, Takamatsu T (2012) Precise analysis of the autofluorescence characteristics of rat colon under UVA and violet light excitation. Curr Pharm Biotechnol (E-pub ahead of print)Google Scholar
- Okada M, Smith NI, Palonpon AF, Endo H, Kawata S, Sodeoka M, Fujita K (2011) Label-free Raman observation of cytochrome c dynamics during apoptosis. Proc Natl Acad Sci USA. doi: 10.1073/pnas.1107524108
- Peres MB, Silveira L Jr, Zangaro RA, Pacheco MT, Pasqualucci CA (2011) Classification model based on Raman spectra of selected morphological and biochemical tissue constituents for identification of atherosclerosis in human coronary arteries. Lasers Med Sci 26(5):645–655. doi: 10.1007/s10103-011-0908-z PubMedCrossRefGoogle Scholar
- Williams PL, Warwick R, Dyson M, Bannister LH (1989) Gray’s anatomy. Churchill Livingstone, LondonGoogle Scholar