Zeiss 2-photon (2P) LSM780 laser scanning confocal microscope

蔡司 2 光子 (2P) LSM780 激光扫描共焦显微镜

• 批准号: 8447387
• 负责人: MOSHE LEVI
• 金额: $ 59.45万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-07-10 至 2014-07-09
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8447387
• 关键词: Area; Atherosclerosis; Biological; Cancer Biology; Collagen; Communities; Complication; DNA; Epithelial; Extracellular Matrix Proteins; Fibrosis; Fluorescence; Fluorescence Microscopy; Fluorescence Resonance Energy Transfer; Fostering; Funding; Generations; Glycolysis; Heart Diseases; Hemorrhage; Image; Kidney Diseases; Label; Lasers; Lipids; Liver diseases; Lung diseases; Malignant Neoplasms; Matrix Metalloproteinases; Medical; Mesenchymal; Metabolism; Microscope; Microscopy; Mitochondria; Modality; Monitor; Oxidative Phosphorylation; Oxidative Stress; Photons; Proteins; Research; Sapphire; Scanning; System; Techniques; Tissues; United States National Institutes of Health; Warburg Effect; base; fluorophore; imaging detector; improved; in vivo; interest; protein protein interaction; second harmonic; spatiotemporal; two-photon

DESCRIPTION (provided by applicant): We are requesting funds to purchase a Carl Zeiss 780 NLO Confocal microscope, which we will couple to a Coherent Chameleon Ultra femto-second Ti: sapphire laser and ISS fast-fluorescence lifetime imaging detector. This system will be the only inverted two-photon microscope generally available at the Anschutz Medical Campus and will be the only fluorescence lifetime imaging system available to the research community in the Greater Denver area. The Zeiss LSM 780 NLO will have tremendous impact on NIH funded research as it is capable of performing Fluorescence Lifetime Imaging Microscopy (FLIM), Two-Photon Excitation (TPE) and Second Harmonic Generation (SHG) Microscopy, and also Hyperspectral Imaging. Fluorescence Lifetime Imaging Microscopy (FLIM): FLIM is becoming a very powerful technique for in vivo spatiotemporal imaging of dynamic changes in mitochondrial metabolism and oxidative stress. In cancer biology the recently reemerging original concept of Warburg effect is also related to alterations in glycolysis versus mitochondrial oxidative phosphorylation. Dynamic imaging of metabolism is achieved in part through monitoring changes in NAD(P)H and FAD lifetimes. FLIM is also a very sensitive technique to perform Foster (or Fluorescence) Resonance Energy Transfer (FRET), a powerful technique to monitor protein-protein interactions of biological interest that occur within a distance of 10 nm or less. Unlike the widely used intensity techniques that are limited by spectral bleed through and spectral overlaps, changes in lifetime can detect real changes in protein-protein interactions without suffering from the limitations associated with the intensity based techniques. Two Photon Excitation (TPE) and Second Harmonic Generation (SHG) Microscopy: SHG is a very powerful technique that allows for label-free imaging of fibrillary collagens. Fibrillary collagens resulting in tissue fibrosis occurs in many pathophysiological conditions associated with tissue fibrosis including kidney disease, liver disease, lung disease, and heart disease. In cancer tissues the so called epithelial to mesenchymal transformation is also associated with accumulation of fibrillary extracellular matrix proteins. In contrast in advanced atherosclerosis activation of tissue matrix metalloproteinases can result in localized decreased in matrix proteins. Spectral Unmixing: This new and improved modality available with the Zeiss LSM780 is a powerful technique that will allow simultaneous imaging of several fluorophores corresponding to several proteins, DNA molecules, or lipids of interest without the complication of bleed through that we have encountered in the past.

描述（由申请人提供）：我们正在申请资金购买 Carl Zeiss 780 NLO 共焦显微镜，我们将其与 Coherent Chameleon Ultra 飞秒钛：蓝宝石激光器和 ISS 快速荧光寿命成像探测器结合使用。该系统将是安舒茨医学园区唯一可用的倒置双光子显微镜，并且将是大丹佛地区研究界可用的唯一荧光寿命成像系统。 Zeiss LSM 780 NLO 将对 NIH 资助的研究产生巨大影响，因为它能够执行荧光寿命成像显微镜 (FLIM)、双光子激发 (TPE) 和二次谐波发生 (SHG) 显微镜以及高光谱成像。荧光寿命成像显微镜 (FLIM)：FLIM 正在成为一种非常强大的技术，用于对线粒体代谢和氧化应激的动态变化进行体内时空成像。在癌症生物学中，最近重新出现的瓦尔堡效应的原始概念也与糖酵解的改变有关 与线粒体氧化磷酸化相比。代谢的动态成像部分是通过监测 NAD(P)H 和 FAD 寿命的变化来实现的。 FLIM 也是一种非常灵敏的技术，可用于执行福斯特（或荧光）共振能量转移 (FRET)，这是一种强大的技术，可监测 10 nm 或更小距离内发生的具有生物意义的蛋白质-蛋白质相互作用。与广泛使用的受光谱渗透和光谱重叠限制的强度技术不同，寿命的变化可以检测蛋白质-蛋白质相互作用的真实变化，而不会受到与基于强度的技术相关的限制。双光子激发 (TPE) 和二次谐波发生 (SHG) 显微镜：SHG 是一种非常强大的技术，可以对纤维胶原蛋白进行无标记成像。导致组织纤维化的纤维胶原发生在与组织纤维化相关的许多病理生理状况中，包括肾病、肝病、肺病和心脏病。在癌症组织中，所谓的上皮至间质转化也与纤维细胞外基质蛋白的积累有关。相反，在晚期动脉粥样硬化中，组织基质金属蛋白酶的激活可导致基质蛋白的局部减少。光谱分离：Zeiss LSM780 提供的这种新的改进模式是一种强大的技术，可以同时对与多种蛋白质、DNA 分子或感兴趣的脂质相对应的多种荧光团进行成像，而不会出现我们过去遇到的渗透并发症。