Complete Fluorescence Spectroscopy Analysis for Oral Cancer Histomorphology

口腔癌組織型態學之完整螢光光譜分析

戴維君(Wei-Chun Tai) ； 陳青淵(Ching Yuan Chen) ； 顏欽堉(Ching-Yu Yen)

《臺灣口腔顎面外科學會雜誌》 29卷1期 (2018/03) Pp. 1-8

摘要

Introduction: Early diagnosis and treatment are important to improve oral cancer patients' survival. Among the early detecting methods, autofluorescence emerged as a potential technology in screening for oral neoplasia. To apply this technique requires a better understanding of the biological basis and complete spectra analysis of optical changes associated with oral cancer. PURPOSE: The goals of this study are to (i) evaluate the broad spectrum of contrast image between cancer and control normal mucosa. (ii) develop available algorithms by analyzing wavelengths to discriminate cancer from normal epithelial for diagnostic utility. MATERIALS AND METHODS: 66 frozen fresh samples were collected from 33 oral cancer patients, (Including 33 tumor and 33 correspond normal). All the samples were collected at the Oral and maxillofacial surgery Department of Chi Mei Medical Center from February 2015 to February 2017. This study was approved by the Institutional Review Board / Ethics Committee (Registration number10401-008). The auto fluorescence spectrum is detected with HITACHI Spectrometer F-7000 high-resolution confocal fluorescence microscopy, the excitation range were set from 250-650nm and emission range were detected from 250 to700 nm. RESULTS: 3 substantial excitation and responding emission intensity peaks between 300 nm and 440 nm (1. 325～335, 2. 355～365 3. 420～440) were found in all the samples. These spectral features agree with the autofluorescence emission pattern of NAD (P) H, Elastin and Flavin. Based on our result, decrease in collagen and elastin contribution was larger than the increase in NADH and Flavin. CONCLUSION: Oral cancer cell shows different consistent, and reflected in autofluorescence intensity compared with the normal mucosa; our premiere result reveals 325～335 nm, 355～365 nm, 420～440 nm excitation wavelength showed more diagnostic capability in oral cancer.

關鍵字

Autofluorescence ； Oral cancer ； Spectrosoopy ； NADH ； Flavin

並列摘要

口腔癌的早期診斷與治療對於病人存活率有重大的影響，在用以早期鑑別診斷工具中，自體螢光被認爲有能早期分辨腫瘤細胞的潛力，但要能有效運用此技術必須要能完整了解整個螢光光譜的結構以及口腔上皮產生病變時所發生的變化。本篇的目標在於(1)分析腫瘤與對照組正常黏膜的廣泛螢光光譜；(2)藉由統計分析來找出兩者之間的差別，進而分辨出光譜中的何種波長最具有鑑別診斷的潛力。本篇實驗對象爲33位確診口腔鱗狀細胞癌的患者，分別由腫瘤處及正常黏膜處留取共66組的檢體，藉由日立F-7000高解析度螢光分析儀進行介析。激發光源波長設定爲250～650奈米，而發射接收波長則設定在250～700奈米。實驗結果發現當以300奈米到440奈米區間中的三種波長(1. 325～335, 2. 355～365, 3. 420～440)激發時可以得到相對應的三個放射波長高峰，這些高峰可能是由細胞自體螢光物質如菸鹼醯胺腺嘌呤二核甘酸，黃素腺鹼二核脢酸，膠原蛋白，彈力蛋白所產生，根據我們的結果膠原蛋白與彈力蛋白的減少影響似乎大於菸鹼醯胺腺嘌呤二核甘酸，黃素腺鹼二核脢酸的增加。結論：口腔癌細胞及正常黏膜細胞在光源照射下發散的自體螢光強度有顯著的差異，結果顯示當予以325～335, 355～365, 420～440奈米波長照射時最具有鑑別診斷的效力。

並列關鍵字

自體螢光 ； 口腔癌 ； 光譜分析 ； 菸鹼醯胺腺嘌呤二核甘酸 ； 黃素腺鹼二核脢酸