Fast super-resolution/confocal microscopy for GI cell biology

用于胃肠道细胞生物学的快速超分辨率/共聚焦显微镜

• 批准号: 10173129
• 负责人: MARK A. MC NIVEN
• 金额: $ 60万
• 依托单位: MAYO CLINIC ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-06-01 至 2022-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10173129
• 关键词: Bioprobe; Biosensor; Cells; Cellular biology; Clinic; Computers; Confocal Microscopy; Coupled; Data Analyses; Detection; Development; Digestive System Disorders; Disease; Dyes; Equipment; Event; Faculty; Fluorescence; Fluorescence Recovery After Photobleaching; Fluorescence Resonance Energy Transfer; Funding; Gastroenterology; Grant; Health; Image; Imaging technology; Individual; Instruction; Lipids; Measures; Microinjections; Microscope; Microscopy; Molecular; Noise; Optics; Pathogenesis; Photobleaching; Photosensitivity; Phototoxicity; Process; Proteins; Research; Research Personnel; Resolution; Services; Signal Pathway; Signal Transduction; Signaling Molecule; Signaling Protein; Speed; Structure; Technology; Three-Dimensional Image; Tissues; Training; United States National Institutes of Health; Work; base; cellular imaging; detector; image reconstruction; imaging modality; improved; innovation; member; new technology; novel; programs; protein protein interaction; recruit; temporal measurement

PROJECT SUMMARY: AIRYSCAN IMAGING TECHNOLOGY The objective of the new equipment is to connect highly productive investigators with state-of-the-art optical technology and applications that facilitate scientific progress and novel breakthroughs. The core user group of well-established cell biologists are members of the NIH-funded Mayo Clinic Center for Cell Signaling in Gas- troenterology (C-SiG). The Specific Aims of the grant are threefold. First, to provide a reliable, accessible, state-of-the-art imaging technology to a user group that will facilitate the study of GI cellular signaling cas- cades. Second, to educate and train individual members of the group in the use of sophisticated cellular imag- ing methods. Emphasis is placed on providing technical instruction and educating investigators on how such approaches can expand the scope and breadth of their independent research programs. Third, to develop and apply innovative fluorescence technologies, including biosensors and vital dyes, to study GI tissues and/or cells in health and disease pathogenesis. This work requires advanced imaging of highly dynamic and/or pho- tosensitive processes within cells while also defining the localization of molecular machinery components with high sensitivity and low phototoxicity. The newly developed Airyscan technology is a major breakthrough that enables this by: i) providing a perfect optical section with highest sensitivity due to its innovative detector array, ii) improving signal-to-noise and spatial resolution to best-in-class, allowing detection of interactions and pro- cesses never before seen, iii) increasing sensitivity to facilitate live imaging of photosensitive processes and structures for long periods (overnight) while minimizing photo damage, and iv) speeding up image acquisition rates for superior temporal resolution to capture highly dynamic events and transient interactions. The new technology will be a part of the C-SiG Optical Microscopy Core, for which confocal microscope utilization has more than tripled in the past 10 years. The Core also provides instruction, technical advice, data interpretation, and development of novel, innovative optical approaches to the study of signaling pathways in GI cells and tis- sues. These services cover a wide range of topics including: confocal microscopy coupled with computer- based 3-D image reconstruction; Fluorescence Resonance Energy Transfer (FRET) applications to measure dynamic protein-protein interactions; Fluorescence Recovery After Photobleaching (FRAP) that allows the quantitation of protein recruitment/turnover; Fluorescence Loss in Photobleaching (FLIP); microinjection of liv- ing cells; expression and use of fluorescence-based bioprobes that facilitates the study and localization of spe- cific signaling proteins and lipids; the development and application of specific photo-activatable caged- compounds that allow a precise temporal and spatial activation of desired signaling molecules in live cells, among others. In addition to supporting a productive NIH-funded user base, prioritized access is provided for promising junior faculty engaged in digestive disease-related research and seeking independent funding.

项目摘要：AIRYS​​CAN 成像技术 新设备的目标是将高生产力的研究人员与最先进的光学连接起来 促进科学进步和新突破的技术和应用。核心用户群为 知名细胞生物学家是美国国立卫生研究院 (NIH) 资助的梅奥诊所气体细胞信号传导中心的成员 肠病学（C-SiG）。该赠款的具体目标有三个。首先，提供可靠、可访问、 向用户群提供最先进的成像技术，这将有助于胃肠道细胞信号传导事件的研究 凯德。其次，教育和培训小组的个别成员使用复杂的细胞成像 荷兰国际集团的方法。重点是提供技术指导并教育调查人员如何进行此类研究 方法可以扩大其独立研究项目的范围和广度。三、发​​展和 应用创新的荧光技术，包括生物传感器和活性染料，来研究胃肠道组织和/或 细胞在健康和疾病发病机制中的作用。这项工作需要高动态和/或光的高级成像 细胞内的敏感过程，同时还定义了分子机械组件的定位 高灵敏度和低光毒性。新开发的 Airyscan 技术是一项重大突破 通过以下方式实现这一点：i) 凭借其创新的探测器阵列提供具有最高灵敏度的完美光学部分， ii) 将信噪比和空间分辨率提高到同类最佳水平，从而能够检测相互作用和亲 以前从未见过的过程，iii）提高灵敏度以促进光敏过程的实时成像和 结构长时间（过夜），同时最大限度地减少照片损坏，以及 iv) 加速图像采集 速率卓越的时间分辨率，以捕获高度动态的事件和瞬态相互作用。新的 技术将成为 C-SiG 光学显微镜核心的一部分，其中共焦显微镜的使用已 过去10年增加了两倍多。核心还提供指导、技术建议、数据解释、 和开发新颖、创新的光学方法来研究胃肠道细胞和组织中的信号通路 起诉。这些服务涵盖广泛的主题，包括： 共焦显微镜与计算机相结合 基于3D图像重建；荧光共振能量转移 (FRET) 应用进行测量 动态蛋白质-蛋白质相互作用；光漂白后的荧光恢复 (FRAP) 允许 蛋白质招募/周转的定量；光漂白中的荧光损失 (FLIP)；活体显微注射 荷兰国际集团细胞；基于荧光的生物探针的表达和使用，有利于特定的研究和定位 特定信号蛋白和脂质；特定光激活笼状材料的开发和应用 允许在活细胞中精确地时间和空间激活所需信号分子的化合物， 除其他外。除了支持 NIH 资助的高效用户群之外，还提供优先访问权 有前途的初级教师从事消化系统疾病相关研究并寻求独立资助。