自體螢光技術已經發展了多年，過去已有研究發現生物體內的腫瘤與正常組織的自體螢光具有不同程度差異，此方法為非侵入式，不需接觸生物體，只需蒐集影像或光譜訊號，已有許多研究應用自體螢光來篩檢癌症，近年來感光元件的技術進步，使相機體積縮小、感光度提高，有研究應用自體螢光影像於體內診斷，甚至有些研究使用相機拍攝口腔中的自體螢光，大範圍拍攝口腔組織，藉此比較病變組織與附近組織的光譜，自體螢光技術已經日益成熟，並且已有應用自體螢光技術的產品在市面上用於輔助醫生篩檢口腔癌，例如：VELscope，在未來應用自體螢光來篩檢癌症必然成為趨勢。
近十年來在台灣口腔癌死亡發生率大幅度提升，患者多數為頰黏膜癌與舌癌，口腔癌前病變發生的早期只要能提早發現並治療，就能降低癌症的發生和死亡，因此早期篩檢口腔癌勢必為重要議題，一般口腔癌篩檢以病理切片檢查確定診斷結果，然而病理切片檢查會造成病人不適，並且檢驗所需時間較長，因此本研究期望應用自體螢光技術開發一種非侵入性篩檢口腔癌影像系統。
生物體內的自體螢光代謝物質主要有還原態菸鹼醯胺腺嘌呤雙核苷酸 (NADH) 以及黃素腺嘌呤二核苷酸 (FAD)，癌症細胞的代謝過程會使NADH的濃度會上升且FAD的濃度會下降。因此本研究將NADH / (NADH + FAD) 稱之為Redox ratio，將其視為代謝狀態。
本研究去年開發了一台能同時獲取NADH及FAD自體螢光的儀器，使其能在臨床試驗使用，但該儀器前端體積較大，頰黏膜部位會較難完整拍攝，因此今年開發一台功能相同且體積較小的儀器，對患者的頰黏膜部位拍攝照片。
在成大醫院牙醫門診挑選口腔癌之患者(確診)、口腔健康之受試者，經過受試者本人同意後，使用兩台口腔螢光偵測儀拍攝其病灶之螢光影像。
分析特徵主要是自體螢光影像之螢光強度、標準差以及Redox ratio的標準差，採用二次分類分析(QDA，Quadratic discriminant analysis)來建立分類器，分別使用去年舊儀器與今年新儀器所收之資料建立分類器及相互測試分類器，研究發現頰黏膜資料所建立的分類器用舌頭資料測試，結果有很高的靈敏度與特異度，研究顯示應用自體螢光影像技術有助於辨識口腔癌。

Current oral cancer screening method is by utilizing tissue biopsy. However, this method is time consuming and causes patients feeling uncomfortable. To overcome this situation, we proposed a non-invasive, real-time method by using autofluorescence image analysis for oral cancer screening system.

Autofluorescence is generated by tissue matrix or fluorophores in living cells. During the period of a tumor developing, the structure of tissue and metabolism will be changed. we are concerned with two fluorescent metabolites, Nicotinamide adenine dinucleotide (NADH) and Flavin adenine dinucleotide (FAD), and its Redox ratio NADH / (NADH +FAD) as our biomarker to indicate the metabolism of tissues.

Last year, we developed a device to observe the NADH/FAD autofluorescence. But this device is so big, it is difficult that take photo of buccal mucosa. Therefore, we developed a smaller device.

we use our device to get clinical data from the Dentistry of National Cheng Kung University Hospital which include the biopsy confirmed oral cancer as the experimental group, and some healthy people are chosen as the control group.

During image analysis, reflectance may interfere with the analysis result. Therefore, we remove reflectance of ROI of autofluorescence image. we analyze the intensity and the heterogeneity of this remove reflectance autofluorescence image and its redox ratio, and use feature for quadratic discriminant analysis (QDA) to create classifier, then use to classify a part data. The testing result shows that its sensitivity and specificity is good.

[1] 李正喆，郭生興，郭英雄，楊博正，韓良俊，頰黏膜癌---最具代表性的檳榔口腔癌，國立台灣大學醫學院附設醫院 牙科部口腔顎面外科，台灣醫學1997年 1卷5期 ,p. 638-641
[2] 梁錦榮，檳榔史暨口腔癌前病變之流行病學，高雄榮民總醫院
[3] 衛生福利部，民國101年主要死因分析，衛生福利部，2012年
[4] 高壽延，陳雅薇，吳政憲，羅文良，雷文天，張哲壽，口腔癌篩檢，台北榮民總醫院
[5] Poate, T., et al., “An audit of the efficacy of the oral brush biopsy technique in a specialist Oral Medicine unit.” Oral oncology, 2004. 40(8): p. 829-834.
[6] Epstein, J., et al., Analysis of oral lesion biopsies identified and evaluated by visual examination, chemiluminescence and toluidine blue. Oral Oncology, 2008. 44(6): p. 538-544.
[7] Farah, C.S., et al., Efficacy of tissue autofluorescence imaging (VELScope) in the visualization of oral mucosal lesions. Head & neck, 2012. 34(6): p. 856-862.
[8] Lingen, M.W., et al., Critical evaluation of diagnostic aids for the detection of oral cancer. Oral oncology, 2008. 44(1): p. 10-22.
[9] Profio, A.E. and D.R. Doiron, A feasibility study of the use of fluorescence bronchoscopy for localization of small lung tumours. Physics in medicine and biology, 1977. 22(5): p. 949.
[10] Bigio, I. J. and J. R. Mourant (1997) Ultraviolet and visible spectroscopy for tissue diagnostics: fluorescence spectroscopy and elastic-scattering spectroscopy. Phys. Med. Bid. 42, 803- 814.
[11] Papazoglou, T. G. (1995) Malignancies and atherosclerotic plaque diagnosis: is laser-induced fluorescence spectroscopy the ultimate solution? J. Photochem. Photobiol. B28, 3-1 I.
[12] Anderson-Engels, S. and B. C. Wilson (1992) In vivo fluorescence in clinical oncology: fundamental and practical issues ,J. Cell. Pharrnacol. 3: p. 66-79.
[13] Richards-Kortum, R. and E. Sevick-Muraca (1996) Quantitative optical spectroscopy for tissue diagnosis. Annu. Rev. Phy,’. Chem. 47: p. 555-606.
[14] Warburg O. The Metabolism of Tumors. London: Arnold Constable, 1930:p.254-270.
[15] 吳玲嘉，李新城，發現癌細胞中能量代謝異常之重要調控因子，生物醫學2012.5(2):p. 88-89
[16] Ostrander, J.H., et al., Optical redox ratio differentiates breast cancer cell lines based on estrogen receptor status. Cancer research, 2010. 70(11): p. 4759-4766.
[17] Drezek, R., et al., Understanding the contributions of NADH and collagen to cervical tissue fluorescence spectra: modeling, measurements, and implications. Journal of biomedical optics, 2001. 6(4): p. 385-396.
[18] Andersson-Engels, S., C. Klinteberg, K. Svanberg and S. Svanberg (1997) In vivo fluorescence imaging for tissue diagnostics. Phys. Med. Biol. 42, 815-824.
[19] D.C.G. De Velda ,M.J.H. Witjes ,H.J.C.M. Sterenborg , J.L.N. Roodenburga, The status of in vivo autofluorescence spectroscopy and imaging for oral oncology , Oral Oncology (2005) 41: p. 117-131
[20] K.H. Awan , P.R. Morgan , S. Warnakulasuriya ,Evaluation of an autofluorescence based imaging system (VELscope™) in the detection of oral potentially malignant disorders and benign keratosis , Oral Oncology 47 (2011): p.274–277.
[21] Farah, C. and M. McCullough, Oral cancer awareness for the general practitioner: new approaches to patient care. Australian dental journal, 2008. 53(1): p. 2-10.
[22] Vander Heiden, M.G., L.C. Cantley, and C.B. Thompson, Understanding the Warburg effect: the metabolic requirements of cell proliferation. science, 2009. 324(5930): p. 1029-1033.
[23] Hsu, P.P. and D.M. Sabatini, Cancer cell metabolism: Warburg and beyond. Cell, 2008. 134(5): p. 703-707.
[24] Liu, Q., et al., Compact point-detection fluorescence spectroscopy system for quantifying intrinsic fluorescence redox ratio in brain cancer diagnostics. Journal of biomedical optics, 2011. 16(3): p. 037004-037004-11.
[25] 陳志魁，應用自體螢光區分正常口腔黏膜、口腔癌前期病變及鱗狀上皮癌，國立陽明大學臨床牙醫學研究所碩士論文，2003年
[26] 陳志魁;江惠華;朱朔嘉;蕭子健;張哲壽，應用自體螢光光譜檢測口腔良性組織及癌前期病灶，中華牙醫學雜誌2004年23卷3期
[27] 蔣維凡，口腔及口咽區域腫瘤解剖生理概論，奇美醫院 ,p1
[28] 潘超群，口腔癌及癌前病變之分類及診斷，台中榮民總醫院
[29] 盧心玉，口腔癌的防治，高雄長庚醫院，聲洋防癌之聲秋季號
[30] 韓良俊，嚼食檳榔行為之預防與戒斷 檳榔之危害―另類物質濫用，台大醫院
[31] 陳嘉瑜，DSP-Based影像擷取系統之開發與研究，國立成功大學工程科學系碩士論文，2004年7月26日
[32] 劉維揚，DSP 影像處理硬體平台之開發，國立中央大學機械工程研究所碩士論文，2001年6月29日
[33] Y. He, N. Khanna, C. J. Boushey and E. J. Delp, Specular highlight removal for image-based dietary assessment, in IEEE International Conference on Multimedia and Expo Workshops, 2012.
[34] Georges A. Wagnieres', Willem M. Star2 and Brian C. Wilson*t3, ln Vivo Fluorescence Spectroscopy and Imaging for Oncological Applications, Photochemistry and Photobiology , 1998. 68(5): p. 603-632,
[35] Kevin Huff, DDS, MAGD , Paul C. Stark, ScD ,Lynn W. Solomon, DDS, MS ,Sensitivity of direct tissue fluorescence visualization in screening for oral premalignant lesions in general practice , Oral Diagnosis General Dentistry ,2009. P.34-38
[36] THOMAS M. COVER ,Geometrical and Statistical Properties of Systems of Linear Inequalities with Applications in Pattern Recognition , IEEE TRANSACTIONS ON ELECTRONIC COMPUTERS ,
[37] Kollias, N., G. Zonios, and G.N. Stamatas, Fluorescence spectroscopy of skin. Vibrational Spectroscopy, 2002. 28(1): p. 17-23.
[38] Melissa Skala, Nirmala Ramanujam, Multiphoton Redox Ratio Imaging for Metabolic Monitoring in vivo, 2010. 594: p. 155–162
[39] Jarvis, R. A., “Focus optimization criteria for computer image processing,” Microscope, 1976. , 24(2): p.163-180.