Tunable White Light Laser Confocal/Spectral Imaging Microscopy

可调谐白光激光共焦/光谱成像显微镜

• 批准号: 7595548
• 负责人: AMMASI PERIASAMY
• 金额: $ 50万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-04-01 至 2010-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7595548
• 关键词: Actins; Alzheimer' s Disease; Area; Behavior; Biological; Cells; Cellular Structures; Cellular biology; Clinical Sciences; Color; Communities; Confocal Microscopy; Country; Disease; Educational workshop; Embryonic Development; Event; Fluorescence Resonance Energy Transfer; Funding; Future; Glutamine; Hand; Housing; Image; Imaging technology; Lasers; Life; Light; Membrane; Microscope; Microscopy; Molecular; Monitor; Non-Invasive Cancer Detection; Pituitary Gland; Process; Protein Dynamics; Proteins; Research; Research Personnel; Sampling; Scientist; System; Therapeutic; Tissues; Training; United States; Universities; Virginia; Walking; biological systems; cellular imaging; design; fluorophore; instrument; interest; light microscopy; protein protein interaction; public health relevance; quantum; research study; sensor

DESCRIPTION (provided by applicant): The W.M. Keck Center for Cellular Imaging (KCCI), an internationally recognized center for advanced light microscopy imaging that is particularly known for protein-protein interaction imaging, is requesting a Leica TCS SP5X "Tunable White Light Laser Confocal/Spectral Imaging Microscopy" system. KCCI is a University wide imaging facility and the requested instrument will benefit the basic and clinical science investigators of this university, and particularly the 8 participating federally funded investigators. The instrument will be housed in the KCCI, located within walking distance of all the participating investigators. The central theme of the research to be performed on the requested Leica TCS SP5X is the study of protein-protein interactions in live cells and other cellular imaging including Photoactivated GFP (PA_GFP) to monitor the dynamic protein interactions in pituitary GHFT1 cells, IQGAPs and the Cell Biology of Alzheimer's Disease, characterization of new FRET pairs, glutamine FRET sensors, study of the mechanism of actin assembly at membranes, and monitoring dynamic behavior of multiple molecular components of the cell biological system during embryonic development. The requested instrument, the Leica SP5X system is unique and represents a quantum leap in confocal microscopy, providing users the ability to tune the excitation and emission wavelengths to match exactly the fluorophores in their experimental samples. The user no longer needs to adapt the experiment to the laser lines available on the confocal system; the tunable white light laser system adapts to the particular experiment. There is currently no other system available at the University of Virginia that has a tunable (470-670 nm) white light laser confocal/spectral imaging capability. This is essential for our core investigator group, and will also be critical for training other investigators interested in the study of protein associations in living cells and tissues. The KCCI is recognized for its annual hands-on-training course, held for the past seven years at the University of Virginia. To date, 304 scientists, from the United States as well as scientists from 23 countries have participated in the KCCI workshop. In summary, the requested instrument will provide the research community at the University of Virginia with a level of imaging capability that is not currently available, and this will enhance significantly ongoing and future research. PUBLIC HEALTH RELEVANCE: The non-invasive detection of biological events inside the living cell was made possible by the many different color fluorescent proteins, but this approach requires a highly adaptable microscope system. The requested Leica TCS SP5X tunable white light laser system is a unique system that permits the user to tune the excitation and emission wavelengths to match exactly the fluorophores in their experimental samples. If we have to understand disease processes and design therapeutic strategies, it is important to continue to develop these live-cell imaging technologies, and the combination of different color fluorescent proteins and the requested Leica SP5X represents a substantial advance in this area.

描述（由申请人提供）：W.M.凯克细胞成像中心 (KCCI) 是国际公认的先进光学显微镜成像中心，尤其以蛋白质-蛋白质相互作用成像而闻名，它正在请求使用徕卡 TCS SP5X“可调谐白光激光共聚焦/光谱成像显微镜”系统。 KCCI 是一个大学范围的成像设施，所需的仪器将使该大学的基础和临床科学研究人员受益，特别是 8 名参与的联邦资助的研究人员。该仪器将安置在 KCCI 内，所有参与研究人员步行即可到达。在所要求的 Leica TCS SP5X 上进行的研究的中心主题是研究活细胞和其他细胞成像中的蛋白质-蛋白质相互作用，包括用于监测垂体 GHFT1 细胞中动态蛋白质相互作用的光激活 GFP (PA_GFP)、IQGAP 和阿尔茨海默病的细胞生物学、新 FRET 对的表征、谷氨酰胺 FRET 传感器、研究 肌动蛋白在膜上的组装，并监测胚胎发育过程中细胞生物系统的多个分子成分的动态行为。所要求的仪器 Leica SP5X 系统是独一无二的，代表了共焦显微镜的巨大飞跃，使用户能够调整激发和发射波长，以精确匹配实验样品中的荧光团。用户不再需要使实验适应共焦系统上可用的激光线；可调谐白光激光系统适应特定的实验。目前，弗吉尼亚大学还没有其他系统具有可调谐（470-670 nm）白光激光共焦/光谱成像功能。这对于我们的核心研究小组至关重要，对于培训对活细胞和组织中蛋白质关联研究感兴趣的其他研究人员也至关重要。 KCCI 因其每年在弗吉尼亚大学举办的实践培训课程而受到认可。迄今为止，已有来自美国的 304 名科学家以及来自 23 个国家的科学家参加了 KCCI 研讨会。总之，所需的仪器将为弗吉尼亚大学的研究界提供目前尚不具备的成像能力，这将显着增强正在进行和未来的研究。公共健康相关性：许多不同颜色的荧光蛋白使活细胞内生物事件的非侵入性检测成为可能，但这种方法需要高度适应性的显微镜系统。所需的 Leica TCS SP5X 可调谐白光激光系统是一个独特的系统，允许用户调整激发和发射波长以精确匹配实验样品中的荧光团。如果我们必须了解疾病过程并设计治疗策略，那么继续开发这些活细胞成像技术非常重要，而不同颜色荧光蛋白与所需的 Leica SP5X 的组合代表了该领域的重大进步。

项目成果

O-Aminobenzoyl-S-nitrosoglutathione: A fluorogenic, cell permeable, pseudo-substrate for S-nitrosoglutathione reductase.

O-氨基苯甲酰基-S-亚硝基谷胱甘肽：一种荧光、细胞可渗透的 S-亚硝基谷胱甘肽还原酶的假底物。

• DOI: 10.1016/j.freeradbiomed.2017.04.008
• 发表时间: 2017
• 期刊: Free radical biology & medicine
• 影响因子: 7.4
• 作者: Sun,BeiLei;Palmer,Lisa;Alam,ShaguftaRehman;Adekoya,Itunuoluwa;Brown-Steinke,Kathleen;Periasamy,Ammasi;Mutus,Bulent
• 通讯作者: Mutus,Bulent

Development of an AP-FRET based analysis for characterizing RNA-protein interactions in myotonic dystrophy (DM1).

• DOI: 10.1371/journal.pone.0095957
• 发表时间: 2014
• 期刊: PloS one
• 影响因子: 3.7
• 作者: Rehman S;Gladman JT;Periasamy A;Sun Y;Mahadevan MS
• 通讯作者: Mahadevan MS

Context-specific regulation of lysosomal lipolysis through network-level diverting of transcription factor interactions

• DOI: 10.1073/pnas.2104832118
• 发表时间: 2021-10
• 期刊: Proceedings of the National Academy of Sciences
• 作者: Vinod K. Mony;Anna Drangowska-Way;R. Albert;E. Harrison;Abbas Ghaddar;M. Horak;Wenfan Ke;Eyleen J. O’Rourke

Scribble mutation disrupts convergent extension and apical constriction during mammalian neural tube closure.

• DOI: 10.1016/j.ydbio.2021.05.013
• 发表时间: 2021-10
• 期刊: Developmental biology
• 影响因子: 2.7
• 作者: Lesko AC;Keller R;Chen P;Sutherland A
• 通讯作者: Sutherland A

Convergent extension in mammalian morphogenesis.

哺乳动物形态发生的趋同延伸。

• DOI: 10.1016/j.semcdb.2019.11.002
• 发表时间: 2020
• 期刊: Seminars in cell & developmental biology
• 影响因子: 7.3
• 作者: Sutherland,Ann;Keller,Raymond;Lesko,Alyssa
• 通讯作者: Lesko,Alyssa