IMAGE NADH AUTOFLUORESCENCE IN MULTICELL TUMOR SPHEROIDS & MONOLAYER CULTURE

以前我们已经证明，细胞自发荧光 当由700-800 nm光激发的多光子主要是由于 NAD（P）H。 在细胞内，这种信号主要来源于 线粒体，其中预计会集中NADH。 研究 在过去的一年里进行的研究试图确定 自发荧光信号，这可以归因于正常的 线粒体功能 两种啮齿动物细胞系（EMT 6，RBL）和一种 迄今为止已经对人细胞系（HeLa）进行了研究。 在这三条线上 细胞自发荧光是动态的，根据状态而变化 的线粒体。 当线粒体解偶联或 等效地，当去除底物时，自发荧光通常 从基础水平下降约20%。 相反，中毒 氰化物的线粒体导致了2.5倍的 信号了 已经确定自发荧光信号是 代谢状态的动态指标，最近的项目已经 利用这种内源性信号作为线粒体健康的报告者。

Previously we have demonstrated that cellular autofluorescence when multiphoton excited by 700-800 nm light is primarily due to NAD(P)H. Within the cell, this signal primarily originates from mitochondria where NADH is expected to be concentrated. Studies conducted within the last year have sought to identify the magnitude of the autofluorescence signal which can be attributed to normal mitochondrial function. Two rodent cell lines (EMT6, RBL) and one human cell line (HeLa) have been examined to date. In all three lines cellular autofluorescence is dynamic, changing according to the status of the mitochondria. When the mitochondria are uncoupled or equivalently when substrate is removed, autofluorescence typically drops by approximately 20% from basal levels. In contrast, poisoning the mitochondria with cyanide results in a 2.5 fold increase in signal. Having established that the autofluorescence signal is a dynamic indicator of metabolic state, more recent projects have utilized this endogenous signal as a reporter of mitochondrial health.