拉曼光譜應用於大腸上皮細胞與癌細胞之分析與診斷能力評估

Analysis and Diagnostic Capability Assessment of Raman Spectrum in Colorectal Cancer and Epithelial Cells

10.6342/NTU201603500

謝承志

大腸癌上皮細胞 ； 拉曼光譜 ； 光譜特徵分析 ； Colorectal epithelial cells ； Raman spectroscopy ； Spectrum feature analysis

臺灣大學生醫電子與資訊學研究所學位論文

2016年

碩士

宋孔彬

繁體中文

本篇主旨為量測大腸上皮細胞與癌細胞之拉曼光譜，再透過軟體將光譜拉曼之外的訊號去除，接著將純粹的拉曼光譜進行細胞基礎物質含量變動的分析，以及每位病人其光譜變動位置差異性。最後再進行一連串的光譜預測，試著探討拉曼光譜在癌症診斷的表現能力是否有機會在之後往臨床發展。 本研究針對大腸上皮細胞(76個)與癌細胞(75個)，共來自七位病人，進行拉曼光譜量測分析。樣本均採用未染色及未封蠟的冷凍切片。分析拉曼結果顯示，每位病人其光譜容易有變動的位置均與整體病人光譜變動位置相符，但有程度大小些許的差異。另外在細胞基本物質組成上面，正常細胞變成癌細胞之後，其DNA組成約上升9%，蛋白質下降了9%，脂質則是大幅度下降到近乎消失的程度。 將拉曼光譜量測結果使用支援向量機(support vector machine)進行leave one out cross validation 預測，得到specificity 100%，sensitivity 88%，整體準確性94%。另外使用leave one patient out cross validation 預測未知病患光譜，得到specificity為76%，sensitivity僅有70%， 整體的準確性為73%。最後單獨將病人自己本身的光譜做leave one out cross validation，整體準確性達98% ，其中specificity為99%，sensitivity為97%。

1. Baraga, J. J., Feld, M. S., & Rava, R. P. (1992). Rapid near-infrared Raman spectroscopy of human tissue with a spectrograph and CCD detector. Applied Spectroscopy, 46(2), 187-190.
   連結：
2. Beljebbar, A., Bouché, O., Diébold, M., Guillou, P., Palot, J., Eudes, D., & Manfait, M. (2009). Identification of Raman spectroscopic markers for the characterization of normal and adenocarcinomatous colonic tissues. Critical reviews in oncology/hematology, 72(3), 255-264.
   連結：
3. Bolzan, M. J. A. (2006). Transformada em ondeleta: Uma necessidade. Revista Brasileira de Ensino de Física, 28(4), 563-567.
   連結：
4. Brennan, J. F., Wang, Y., Dasari, R. R., & Feld, M. S. (1997). Near-infrared Raman spectrometer systems for human tissue studies. Applied Spectroscopy, 51(2), 201-208.
   連結：
5. Chang, C.-C., & Lin, C.-J. (2011). LIBSVM: a library for support vector machines. ACM Transactions on Intelligent Systems and Technology (TIST), 2(3), 27.
   連結：
6. Duraipandian, S., Bergholt, M. S., Zheng, W., Ho, K. Y., Teh, M., Yeoh, K. G., . . . Huang, Z. (2012). Real-time Raman spectroscopy for in vivo, online gastric cancer diagnosis during clinical endoscopic examination. Journal of biomedical optics, 17(8), 081418-081418.
   連結：
7. Duraipandian, S., Zheng, W., Ng, J., Low, J. J., Ilancheran, A., & Huang, Z. (2012). Simultaneous fingerprint and high-wavenumber confocal Raman spectroscopy enhances early detection of cervical precancer in vivo. Analytical chemistry, 84(14), 5913-5919.
   連結：
8. Han, J., Pei, J., & Kamber, M. (2011). Data mining: concepts and techniques: Elsevier.
   連結：
9. Lieber, C. A., & Mahadevan-Jansen, A. (2003). Automated method for subtraction of fluorescence from biological Raman spectra. Applied Spectroscopy, 57(11), 1363-1367.
   連結：
10. Lue, N., Kang, J. W., Yu, C.-C., Barman, I., Dingari, N. C., Feld, M. S., . . . Fitzmaurice, M. (2012). Portable optical fiber probe-based spectroscopic scanner for rapid cancer diagnosis: a new tool for intraoperative margin assessment. PloS one, 7(1), e30887.
    連結：
11. Lui, H., Zhao, J., McLean, D., & Zeng, H. (2012). Real-time Raman spectroscopy for in vivo skin cancer diagnosis. Cancer research, 72(10), 2491-2500.
    連結：
12. Lyng, F. M., Faoláin, E. Ó., Conroy, J., Meade, A., Knief, P., Duffy, B., . . . Byrne, H. (2007). Vibrational spectroscopy for cervical cancer pathology, from biochemical analysis to diagnostic tool. Experimental and molecular pathology, 82(2), 121-129.
    連結：
13. Mahadevan-Jansen, A., & Richards-Kortum, R. R. (1996). Raman spectroscopy for the detection of cancers and precancers. Journal of biomedical optics, 1(1), 31-70.
    連結：
14. McCreery, R. L. (2005). Raman spectroscopy for chemical analysis (Vol. 225): John Wiley & Sons.
    連結：
15. Mircea Mujat, Kristyn Greco, Kristin L. Galbally-Kinney, Daniel X. Hammer, R. Daniel Ferguson, Nicusor Iftimia, Phillip Mulhall, Puneet Sharma, Michael J. Pikal, and William J. Kessler. (2012). Optical coherence tomography-based freeze-drying microscopy. Biomed Opt Express.
    連結：
16. Morris, M. D., Matousek, P., Towrie, M., Parker, A. W., Goodship, A. E., & Draper, E. R. (2005). Kerr-gated time-resolved Raman spectroscopy of equine cortical bone tissue. Journal of biomedical optics, 10(1), 014014-0140147.
    連結：
17. Movasaghi, Z., Rehman, S., & Rehman, I. U. (2007). Raman spectroscopy of biological tissues. Applied Spectroscopy Reviews, 42(5), 493-541.
    連結：
18. Nijssen, A., Koljenović, S., Schut, T. C. B., Caspers, P. J., & Puppels, G. J. (2009). Towards oncological application of Raman spectroscopy. Journal of biophotonics, 2(1‐2), 29-36.
    連結：
19. Raman, C. (1929). Part II.—The Raman effect. Investigation of molecular structure by light scattering. Transactions of the Faraday Society, 25, 781-792.
    連結：
20. Ramos, P. M., & Ruisánchez, I. (2005). Noise and background removal in Raman spectra of ancient pigments using wavelet transform. Journal of Raman Spectroscopy, 36(9), 848-856.
    連結：
21. Robichaux-Viehoever, A., Kanter, E., Shappell, H., Billheimer, D., Jones, H., & Mahadevan-Jansen, A. (2007). Characterization of Raman spectra measured in vivo for the detection of cervical dysplasia. Applied Spectroscopy, 61(9), 986-993.
    連結：
22. Shlens, J. (2014). A tutorial on principal component analysis. arXiv preprint arXiv:1404.1100.
    連結：
23. Short, M. A., Lam, S., McWilliams, A. M., Ionescu, D. N., & Zeng, H. (2011). Using laser Raman spectroscopy to reduce false positives of autofluorescence bronchoscopies: a pilot study. Journal of Thoracic Oncology, 6(7), 1206-1214.
    連結：
24. Shreve, A. P., Cherepy, N. J., & Mathies, R. A. (1992). Effective rejection of fluorescence interference in Raman spectroscopy using a shifted excitation difference technique. Applied Spectroscopy, 46(4), 707-711.
    連結：
25. Silveira Jr, L., Bodanese, B., Zangaro, R. A., & Pacheco, M. T. T. (2010). Discrete wavelet transform for denoising Raman spectra of human skin tissues used in a discriminant diagnostic algorithm. Instrumentation Science & Technology, 38(4), 268-282.
    連結：
26. Szymanski, H. A. (2012). Raman spectroscopy: theory and practice: Springer Science & Business Media.
    連結：
27. Zhao, J., Lui, H., McLean, D. I., & Zeng, H. (2007). Automated autofluorescence background subtraction algorithm for biomedical Raman spectroscopy. Applied Spectroscopy, 61(11), 1225-1232.
    連結：
28. Hricak, H., Choi, B. I., Scott, A. M., Sugimura, K., Muellner, A., von Schulthess, G. K., . . . Larson, S. M. (2010). Global trends in hybrid imaging 1. Radiology, 257(2), 498-506.
29. Jermyn, M., Mok, K., Mercier, J., Desroches, J., Pichette, J., Saint-Arnaud, K., . . . Leblond, F. (2015). Intraoperative brain cancer detection with Raman spectroscopy in humans. Science translational medicine, 7(274), 274ra219-274ra219.
30. Pang-Ning, T., Steinbach, M., & Kumar, V. (2006). Introduction to data mining. Paper presented at the Library of congress.