Colon Adenocarcinoma Diagnosis in Human Samples by Multicontrast Nonlinear Optical Microscopy of Hematoxylin and Eosin Stained Histological Sections

Javier Adur^1,2,3, Mariana Bianchi¹, Vitor B. Pelegati³, Silvia Viale¹, María F. Izaguirre¹, Hernandes F. Carvalho^3,4, Carlos L. Cesar^4,5, Víctor H. Casco^1*

¹Microscopy Laboratory Applied to Molecular and Cellular Studies, School of Engineering, National University of Entre Ríos (UNER), Oro Verde, Argentina.
²Research and Transfer Center of Entre Ríos (CITER) CONICET-UNER, Oro Verde, Argentina.
³INFABiC-National Institute of Science and Technology on Photonics Applied to Cell Biology, Campinas, Brazil.
⁴Department of Structural and Functional Biology, Institute of Biology, State University of Campinas (UNICAMP), Campinas, Brazil.
⁵Biophotonic Group, Optics and Photonics Research Center (CEPOF), Institute of Physics “Gleb Wataghin”, State University of Campinas (UNICAMP), Campinas, Brazil.

DOI: 10.4236/jct.2014.513127                

Abstract

Combined multimodal nonlinear optical (NLO) microscopies were used to detect and quantify morphological changes associated with stroma and epithelial transformation in colon cancer. Our findings provide complementary information about tissue microstructure, displaying distinctive patterns between normal and malignant human colon. Additionally, we have demonstrated the usefulness of using fixed tissues for the disease diagnostic and prognostic. The present work provides a framework for using NLO techniques as a clinical diagnostic tool for human colon cancer. NLO metrics could be applied to other disorders, which are characterized by abnormal cell proliferation and collagen assembly.

Keywords

Nonlinear Microscopy, Nonlinear Multicontrast, Histological Sections, Human Colon Cancer

Conflicts of Interest

The authors declare no conflicts of interest.

References

[1]	Jemal, A., Bray, F., Center, M.M., Ferlay, J., Ward, E. and Forman, D. (2011) Global Cancer Statistics. CA: A Cancer Journal for Clinicians, 61, 69-90. http://dx.doi.org/10.3322/caac.20107
[2]	Taylor, J.C., Kendall, C.A., Stone, N. and Cook, T.A. (2007) Optical Adjuncts for Enhanced Colonoscopic Diagnosis. British Journal of Surgery, 94, 6-16. http://dx.doi.org/10.1002/bjs.5628
[3]	Rex, D.K. (2007) Colonoscopy: The Dominant and Preferred Colorectal Cancer Screening Strategy in the United States. Mayo Clinic Proceedings, 82, 662-664. http://dx.doi.org/10.1016/S0025-6196(11)61183-X
[4]	Lopes, P.C., Moreira, J.A., Almeida, A., Esteves, A., Gregora, I., Ledinsky, M., Lopes, J.M., Henrique, R. and Oliveira, A. (2011) Discriminating Adenocarcinoma from Normal Colonic Mucosa through Deconvolution of Raman Spectra. Journal of Biomedical Optics, 16, Article ID: 127001.
[5]	Banerjee, B., Renkoski, T., Graves, L.R., Rial, N.S., Tsikitis, V.L., Nfonsam, V., Pugh, J., Tiwari, P., Gavini, H. and Utzinger, U. (2012) Tryptophan Autofluorescence Imaging of Neoplasms of the Human Colon. Journal of Biomedical Optics, 17, Article ID: 016003. http://dx.doi.org/10.1117/1.JBO.17.1.016003
[6]	Imaizumi, K., Harada, Y., Wakabayashi, N., Yamaoka, Y., Konishi, H., Dai, P., Tanaka, H. and Takamatsu, T. (2012) Dual-Wavelength Excitation of Mucosal Autofluorescence for Precise Detection of Diminutive Colonic Adenomas. Gastrointestinal Endoscopy, 75, 110-117. http://dx.doi.org/10.1016/j.gie.2011.08.012
[7]	Cicchi, R., Sturiale, A., Nesi, G., Tonelli, F. and Pavone, F.S. (2012) Two-Photon Imaging and Spectroscopy of Fresh Human Colon Biopsies. Proceedings of SPIE, 8226. http://dx.doi.org/10.1117/12.907822
[8]	Tadrous, P.J. (2000) Methods for Imaging the Structure and Function of Living Tissues and Cells: Fluorescence Lifetime Imaging. Journal of Pathology, 191, 229-234. http://dx.doi.org/10.1002/1096-9896(200007)191:3<229::AID-PATH623>3.0.CO;2-B
[9]	Tadrous, P.J., Siegel, J., French, P.M., Shousha, S., Lalani, E.-N. and Stamp, G.W. (2003) Fluorescence Lifetime Imaging of Unstained Tissues: Early Results in Human Breast Cancer. Journal of Pathology, 199, 309-317. http://dx.doi.org/10.1002/path.1286
[10]	Carriles, R., Schafer, D.N., Sheetz, K.E., Field, J.J., Cisek, R., Barzda, V., Sylvester, A.W. and Squier, J.A. (2009) Invited Review Article: Imaging Techniques for Harmonic and Multiphoton Absorption Fluorescence Microscopy. Review of Scientific Instruments, 8, Article ID: 081101.
[11]	Campagnola, P.J. and Dong, C.Y. (2011) Second Harmonic Generation Microscopy: Principles and Applications to Disease Diagnosis. Laser & Photonics Reviews, 5, 13-26. http://dx.doi.org/10.1002/lpor.200910024
[12]	Zhuo, S., Zhu, X., Wu, G., Chen, J. and Xie, S. (2011) Quantitative Biomarkers of Colonic Dysplasia Based on Intrinsic Second-Harmonic Generation Signal. Journal of Biomedical Optics, 16, Article ID: 120501. http://dx.doi.org/10.1117/1.3659715
[13]	Débarre, D., Supatto, W., Pena, A.M., Fabre, A., Tordjmann, T., Combettes, L., Schanne-Klein, M.C. and Beaurepaire, E. (2006) Imaging Lipid Bodies in Cells and Tissues Using Third-Harmonic Generation Microscopy. Nature Methods, 3, 47-53. http://dx.doi.org/10.1038/nmeth813
[14]	Zipfel, W.R., Williams, R.M. and Webb, W.W. (2003) Nonlinear Magic: Multiphoton Microscopy in the Biosciences. Nature Biotechnology, 21, 1369-1377. http://dx.doi.org/10.1038/nbt899
[15]	Cicchi, R., Massi, D., Sestini, S., Carli, P., De Giorgi, V., Lotti, T. and Pavone, F.S. (2007) Multidimensional Non-Linear Laser Imaging of Basal Cell Carcinoma. Optics Express, 15, 10135-10148. http://dx.doi.org/10.1364/OE.15.010135
[16]	Tuer, A., Tokarz, D., Prent, N., Cisek, R., Alami, J., Dumont, D.J., Bakueva, L., Rowlands, J. and Barzda, V. (2010) Nonlinear Multicontrast Microscopy of Hematoxylin-and-Eosin-Stained Histological Sections. Journal of Biomedical Optics, 15, Article ID: 026018. http://dx.doi.org/10.1117/1.3382908
[17]	Adur, J., Pelegati, V.B., de Thomaz, A.A., Baratti, M.O., Almeida, D.B., Andrade, L.A., Bottcher-Luiz, F., Carvalho, H.F. and Cesar, C.L. (2012) Optical Biomarkers of Serous and Mucinous Human Ovarian Tumor Assessed with Nonlinear Optics Microscopies. PLoS ONE, 7, e47007. http://dx.doi.org/10.1371/journal.pone.0047007
[18]	Adur, J., Pelegati, V.B., Costa, L.F., Pietro, L., de Thomaz, A.A., Almeida, D.B., Bottcher-Luiz, F., Andrade, L.A. and Cesar, C.L. (2011) Recognition of Serous Ovarian Tumors in Human Samples by Multimodal Nonlinear Optical Microscopy. Journal of Biomedical Optics, 16, Article ID: 096017. http://dx.doi.org/10.1117/1.3626575
[19]	Adur, J., Pelegati, V.B., de Thomaz, A.A., D’Souza-Li, L., Assun??o Mdo, C., Bottcher-Luiz, F., Andrade, L.A. and Cesar, C.L. (2012) Quantitative Changes in Human Epithelial Cancers and Osteogenesis Imperfecta Disease Detected Using Nonlinear Multicontrast Microscopy. Journal of Biomedical Optics, 17, Article ID: 081407. http://dx.doi.org/10.1117/1.JBO.17.8.081407
[20]	Conklin, M.W., Eickhoff, J.C., Riching, K.M., Pehlke, C.A., Eliceiri, K.W., Provenzano, P.P., Friedl, A. and Keely, P.J. (2011) Aligned Collagen Is a Prognostic Signature for Survival in Human Breast Carcinoma. The American Journal of Pathology, 178, 1221-1232. http://dx.doi.org/10.1016/j.ajpath.2010.11.076
[21]	Provenzano, P.P., Eliceiri, K.W., Campbell, J.M., Inman, D.R., White, J.G. and Keely, P.J. (2006) Collagen Reorganization at the Tumor-Stromal Interface Facilitates Local Invasion. BMC Medicine, 4, 38. http://dx.doi.org/10.1186/1741-7015-4-38
[22]	Yu, C.H., Tai, S.P., Kung, C.T., Lee, W.J., Chan, Y.F., Liu, H.L., Lyu, J.Y. and Sun, C.K. (2008) Molecular Third-Harmonic-Generation Microscopy through Resonance Enhancement with Absorbing Dye. Optics Letters, 33, 387-389. http://dx.doi.org/10.1364/OL.33.000387
[23]	De Carvalho, H.F. and Taboga, S.R. (1996) The Applicability of Hematoxylin-Eosin Staining plus Fluorescence or Confocal Laser Scanning Microscopy to the Study of Elastic Fibers in Cartilages. Comptes Rendus de l’Académie des Sciences—Series III—Sciences de la Vie, 319, 991-996.
[24]	Drezek, R., Brookner, C., Pavlova, I., Boiko, I., Malpica, A., Lotan, R., Follen, M. and Richards-Kortum, R. (2001) Autofluorescence Microscopy of Fresh Cervical-Tissue Sections Reveals Alterations in Tissue Biochemistry with Dysplasia. Photochemistry and Photobiology, 73, 636-641. http://dx.doi.org/10.1562/0031-8655(2001)0730636AMOFCT2.0.CO2
[25]	Bird, D.K., Yan, L., Vrotsos, K.M., Eliceiri, K.W., Vaughan, E.M., Keely, P.J., White, J.G. and Ramanujam, N. (2005) Metabolic Mapping of MCF10A Human Breast Cells via Multiphoton Fluorescence Lifetime Imaging of the Coenzyme NADH. Cancer Research, 65, 8766-8773. http://dx.doi.org/10.1158/0008-5472.CAN-04-3922
[26]	Provenzano, P.P., Rueden, C.T., Trier, S.M., Yan, L., Ponik, S.M., Inman, D.R., Keely, P.J. and Eliceiri, K.W. (2008) Nonlinear Optical Imaging and Spectral-Lifetime Computational Analysis of Endogenous and Exogenous Fluorophores in Breast Cancer. Journal of Biomedical Optics, 13, Article ID: 031220. http://dx.doi.org/10.1117/1.2940365
[27]	Cicchi, R., Sturiale, A., Nesi, G., Kapsokalyvas, D., Alemanno, G., Tonelli, F. and Pavone, F.S. (2013) Multiphoton Morpho-Functional Imaging of Healthy Colon Mucosa, Adenomatous Polyp and Adenocarcinoma. Biomedical Optics Express, 4, 1204-1213. http://dx.doi.org/10.1364/BOE.4.00120432
[28]	Birk, J.W., Tadros, M., Moezardalan, K., Nadyarnykh, O., Forouhar, F., Anderson, J. and Campagnola, P. (2014) Second Harmonic Generation Imaging Distinguishes Both High-Grade Dysplasia and Cancer from Normal Colonic Mucosa. Digestive Diseases and Sciences, 59, 1529-1534.