In vivo multiphoton fluorescence lifetime imaging of protein-bound and free nicotinamide adenine dinucleotide in normal and precancerous epithelia
In vivo multiphoton fluorescence lifetime imaging of protein-bound and free nicotinamide adenine dinucleotide in normal and precancerous epithelia
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DOI：
10.1117/1.2717503
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发表时间：
2007-03-01
影响因子：
3.5
通讯作者：
Ramanujam, Nirmala
中科院分区：
文献类型：
--
作者：
Skala, Melissa C.;Riching, Kristin M.;Ramanujam, Nirmala
多光子荧光寿命成像显微镜 (FLIM) 是一种无创、细胞分辨率、3D 功能成像技术。我们研究了通过内源性代谢辅因子烟酰胺腺嘌呤二核苷酸 (NADH) 的多光子 FLIM 进行代谢成像进行体内癌前诊断的潜力。使用多光子 FLIM 在 780 nm 激发下对经二甲基苯并 α 蒽 (DMBA) 处理的仓鼠颊囊口腔癌模型和 MCF10A 细胞单层进行成像。正常仓鼠颊囊上皮细胞的细胞质具有短寿命（0.29 +/- 0.03 ns）和长寿命成分（2.03 +/- 0.06 ns），分别归因于游离和蛋白质结合的 NADH。低度癌前病变（轻度至中度不典型增生）和高度癌前病变（重度不典型增生和原位癌）与正常组织的区别在于其蛋白质结合的 NADH 寿命缩短（p < 0.05）。细胞单层细胞糖酵解和氧化磷酸化的抑制分别导致蛋白质结合的 NADH 寿命增加和减少 (p < 0.05)。结果表明，不典型增生导致蛋白质结合的 NADH 寿命缩短是由于从氧化磷酸化转变为糖酵解，这与肿瘤代谢的预测一致。我们证明多光子 FLIM 是一种强大的工具，用于体内上皮癌前病变的无创表征和检测。 (C) 2007 年光电仪器工程师协会。
Multiphoton fluorescence lifetime imaging microscopy (FLIM) is a noninvasive, cellular resolution, 3-D functional imaging technique. We investigate the potential for in vivo precancer diagnosis with metabolic imaging via multiphoton FLIM of the endogenous metabolic cofactor nicotinamide adenine dinucleotide (NADH). The dimethylbenz [alpha] anthracene (DMBA)-treated hamster cheek pouch model of oral carcinogenesis and MCF10A cell monolayers are imaged using multiphoton FLIM at 780-nm excitation. The cytoplasm of normal hamster cheek pouch epithelial cells has short (0.29 +/- 0.03 ns) and long lifetime components (2.03 +/- 0.06 ns), attributed to free and protein-bound NADH, respectively. Low-grade precancers (mild to moderate dysplasia) and high-grade precancers (severe dysplasia and carcinoma in situ) are discriminated from normal tissues by their decreased protein-bound NADH lifetime (p < 0.05). Inhibition of cellular glycolysis and oxidative phosphorylation in cell monolayers produces an increase and decrease, respectively, in the protein-bound NADH lifetime (p < 0.05). Results indicate that the decrease in protein-bound NADH lifetime with dysplasia is due to a shift from oxidative phosphorylation to glycolysis, consistent with the predictions of neoplastic metabolism. We demonstrate that multiphoton FLIM is a powerful tool for the noninvasive characterization and detection of epithelial precancers in vivo. (C) 2007 Society of Photo-Optical Instrumentation Engineers.