FLIM as a tool for metabolic imaging of the cornea

A. Batista, C. Loureiro, J. Domingues, J. S. Silva, M. Morgado
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Abstract

We intend to develop an efficient method of measuring respiratory function of the cornea. With this purpose, we resorted to fluorescence lifetime imaging microscopy (FLIM) to monitor the metabolic co-factor flavin adenine dinucleotide (FAD). FAD and nicotinamide adenine dinucleotide (NADH) are co-factors of the electron transport chain. Therefore alterations in amount of these molecules reflect alterations in the metabolism. For assessing the potential of FLIM for metabolic imaging of the cornea, we performed a series of experiments using a time-correlated single photon counting (TCSPC) fluorescence lifetime microscope (Picoquant to MicroTime 100 coupled to an Olympus BX51 Microscope). In this technique, the acquired signal is the convolution between the instrument response function (IRF) and the fluorescence signal from the sample. IRF was acquired using an Erythrosin B solution. In this work we show that it is possible to acquire fluorescence lifetime images of rat and bovine corneas using FAD autofluorescence.
FLIM作为角膜代谢成像的工具
我们打算发展一种有效的测量角膜呼吸功能的方法。为此,我们采用荧光寿命成像显微镜(FLIM)监测代谢辅助因子黄素腺嘌呤二核苷酸(FAD)。FAD和烟酰胺腺嘌呤二核苷酸(NADH)是电子传递链的辅助因子。因此,这些分子数量的变化反映了代谢的变化。为了评估FLIM在角膜代谢成像中的潜力,我们使用时间相关单光子计数(TCSPC)荧光寿命显微镜(Picoquant to MicroTime 100与Olympus BX51显微镜耦合)进行了一系列实验。在该技术中,采集的信号是仪器响应函数(IRF)与样品荧光信号之间的卷积。IRF用红素B溶液获得。在这项工作中,我们表明,它是可能获得荧光寿命图像的大鼠和牛角膜使用FAD自体荧光。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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