MITOCHONDRIAL REDOX IMAGING OF BIOLOGICAL TISSUES

这个子项目是许多利用资源的研究子项目之一 由NIH/NCRR资助的中心拨款提供。子项目的主要支持 而子项目的主要调查员可能是由其他来源提供的， 包括其它NIH来源。 列出的子项目总成本可能 代表子项目使用的中心基础设施的估计数量， 而不是由NCRR赠款提供给子项目或子项目工作人员的直接资金。 用于成像线粒体代谢功能的尖端技术的发展 包括生物组织的氧化还原状态的研究是非常需要的，因为线粒体及其 功能/遗传异常与许多疾病包括癌症有关， 糖尿病、心血管疾病和神经退行性疾病。低温NADH/Fp （还原型烟酰胺腺嘌呤二核苷酸/氧化型黄素蛋白）荧光成像或 自大约30年前以来，已经开发了“氧化还原扫描”来对体内 基于氧化还原比率（Fp/NADH或Fp/（NADH+Fp）），测定组织的线粒体氧化还原状态。 还建立了基于CCD的氧化还原成像仪，以更快地获取图像，具有更高的空间分辨率。 分辨率（~10 pm）。然而，氧化还原比是基于氧化还原的相对强度的比率。 NADH和Fp的荧光信号，这通常取决于仪器设置， 过滤器和灯的条件。在这个项目中，我们的目标是开发一个校准程序，以量化 NADH和Fp信号，以便在不同时间或不同设置下获得氧化还原图像 可以比较。我们还旨在升级旧氧化还原扫描仪的硬件和软件 到一个新的pc控制单元，将获得氧化还原图像更快，更高的信噪比和信号 动态范围 我们还致力于开发活性氧的组织成像方法， 它们与线粒体的氧化还原状态有关，也被认为在细胞凋亡中起关键作用。 许多疾病的发生和发展。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. The development of cutting edge technologies for imaging the mitochondrial metabolic function including the redox state of biological tissues are in great need, since mitochondria and its functional/genetic abnormalities have been connected to a number of diseases including cancer, diabetes, cardiovascular diseases, and neurodegenerative diseases. Low temperature NADH/Fp (reduced nicotinamide adenine dinucleotide/oxidized flavoproteins) fluorescence imaging or "redox scanning" has been developed since about 30 years ago to image the in vivo mitochondrial redox states of tissues on the basis of the redox ratios (Fp/NADH or Fp/(NADH+Fp)). A CCD-based redox imager has also been built to acquire images much faster with higher spatial resolution (~10 pm). However, redox ratio is based on the ratio of the relative intensities of the fluorescence signals of NADH and Fp, which usually depend on instrument settings such as filters and lamp conditions. In this project we aim to develop a calibration procedure to quantify NADH and Fp signals so that redox images obtained at different time or with different settings can be compared. We also aim to upgrade the hardware and software of the old redox scanner to a new pc-control unit that will acquire redox images faster with higher SNR and signal dynamic ranges. We also aim to develop tissue imaging methods for reactive oxygen species, which are connected to mitochondrial redox state and also implicated to play key role in the generation and progression of many diseases.