Acquisition of Zeiss LSM980 with Airyscan 2, a super-resolution point scanning confocal microscope

购买 Zeiss LSM980 和 Airyscan 2（超分辨率点扫描共焦显微镜）

• 批准号: 10632893
• 负责人: RANDAL J. KAUFMAN
• 金额: $ 60万
• 依托单位: SANFORD BURNHAM PREBYS MEDICAL DISCOVERY INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-06-05 至 2024-06-04
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10632893
• 关键词: Aging; Binding; Cells; Collaborations; Communities; Data; Development; Disease; Drosophila genus; Dyes; Embryo; Epigenetic Process; Event; Funding; Homeostasis; Image; Joints; Kinetics; Location; Malignant Neoplasms; Metabolism; Microscope; Molecular; Nerve Degeneration; Noise; Organoids; Physiological; Resolution; Sampling; Scanning; Scientific Advances and Accomplishments; Slice; Speed; System; Technology; Thick; Time; Tissue imaging; Tissues; Visualization; age related; detector; equipment acquisition; fluorophore; grasp; interest; novel; optical spectra; quasar; stem cell biology; superresolution imaging; superresolution microscopy; temporal measurement; time use; trafficking; ultra high resolution

PROJECT SUMMARY Sanford Burnham Prebys (SBP) hosts labs with a very broad range of interests which include cancer, aging and age-related diseases like neurodegeneration, cell and tissue homeostasis and function, metabolism and stem cell biology. We truly believe that visualization of molecules of interest, using dyes or fluorophores, in the context of both their spatial and temporal location within cells or tissues is key to understanding physiological attributes of normal versus diseased states. Super-resolution microscopy has become a key technology in providing this context. However, an increase in spatial resolving power often requires a sacrifice in temporal resolution i.e. the speed at which molecular kinetics of fluorophores can be observed, which necessitates careful selection of the super-resolution mode required to fulfill experimental needs. For this purpose, SBP requests support for a Zeiss LSM980 with Airyscan 2. The LSM980 is capable of simultaneously imaging up to 4 different fluorophores with well-separated, non-overlapping emission spectra with a two-fold increase in spatial resolution. The Multiplex modes, only available on the LSM980, can speed up image acquisition up to twenty five-fold beyond acquisition rates on typical point-scanning confocal microscopes, and still provide a near two-fold increase in spatial resolution. This speed is essential in studying highly dynamic mechanisms that require higher spatial resolution, such as vesicular trafficking, transient molecular interactions or epigenetic changes. The ability to gauge subtle differences in these highly dynamic events will give our users a better grasp of molecular changes that occur from healthy to diseased systems. This speed of the Multiplex modes is also essential for fixed thick tissue imaging (>200µm) where large fields of view or thick slices can be imaged in a fraction of the time, at super-resolution. The LSM980 brings additional imaging benefits not currently available at SBP. The Quasar detector permits simultaneous imaging of the full emission spectrum i.e. all fluorophores. The LSM980 is more sensitive to weak samples and less sensitive to background noise, and the newly added Airyscan joint deconvolution processing provides 90nm spatial resolution. These features make it essential for projects requiring super-resolution imaging of thick samples, for e.g. organoids, cell clusters or drosophila embryos. As the LSM980 will be the first available for the San Diego community, the scientific advances made possible by acquisition of this instrument at SBP will allow the 12 major users with 75% of the accessible usage time (AUT) to generate previously unattainable data, which will further development of novel hypothesis, new funding and both internal and external collaborations.

项目摘要 Sanford Burnham Prebys（SBP）主持具有非常广泛兴趣的实验室，包括 癌症，衰老和与年龄相关的疾病，例如神经退行性，细胞和组织稳态以及 功能，代谢和干细胞生物学。我们真正相信的可视化分子的可视化 在其空间和临时位置的背景下，使用染料或荧光团的兴趣 细胞或组织是理解正常状态与患病状态的物理属性的关键。 超分辨率显微镜已成为提供此环境的关键技术。然而， 空间分辨率的增加通常需要暂时的分辨率牺牲，即 可以观察到荧光团的分子动力学的速度，必要 选择满足实验需求所需的超分辨率模式。为此， SBP请求支持Zeiss LSM980与Airyscan 2。LSM980能够 类似地，具有多达4种不同的荧光团的成像 发射光谱，空间分辨率增加了两倍。多重模式，仅 在LSM980上可用，可以加快图像获取的速度超出25倍 典型的扫描共聚焦显微镜的获取率，但仍提供近两个倍 空间分辨率的增加。这种速度对于研究高度动态机制至关重要 需要更高的空间分辨率，例如囊泡运输，瞬态分子相互作用或 表观遗传变化。在这些高度动态事件中衡量微妙差异的能力将 使我们的用户更好地了解从健康到脱离系统发生的分子变化。 多路复用模式的这种速度对于固定厚的组织成像（> 200µm）也至关重要 可以在超级分辨率的一小部分中成像大的视野或厚切片。这 LSM980带来了SBP目前尚未获得的其他成像好处。类星体检测器 允许同时进行全发射光谱，即所有荧光团。 LSM980是 对弱样品更敏感，对背景噪声敏感，新添加 Airyscan关节反卷积处理可提供90NM的空间分辨率。这些功能构成 对于需要厚样品的超分辨率成像的项目至关重要，例如器官， 细胞簇或Dropophila胚胎。因为LSM980将是圣地亚哥的第一个 社区，科学进步通过在SBP中获取该工具而成为可能 允许12个具有75％可访问用法时间（AUT）的主要用户以前生成 无法实现的数据，这将进一步发展新的假设，新资金以及两者 内部和外部协作。