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.

项目概要 桑福德伯纳姆普雷比学院 (SBP) 拥有兴趣广泛的实验室，其中包括 癌症、衰老和与年龄相关的疾病，如神经退行性变、细胞和组织稳态以及 我们坚信分子的可视化。 兴趣，使用染料或荧光团，在其空间和时间位置的背景下 细胞或组织是理解正常状态与疾病状态的生理属性的关键。 超分辨率显微镜已成为提供这种背景的关键技术。 空间分辨率的提高通常需要牺牲时间分辨率，即 观察荧光团分子动力学的速度，这需要仔细 选择满足实验需求所需的超分辨率模式。 SBP 请求支持带有 Airyscan 2 的 Zeiss LSM980。LSM980 能够 同时对多达 4 个不同的荧光团进行充分分离、不重叠的成像 仅在多重模式下，发射光谱的空间分辨率提高了两倍。 LSM980 上可用，可将图像采集速度提高 25 倍 典型点扫描共焦显微镜的采集率，并且仍然提供近两倍的采集率 空间分辨率的提高对于研究高动态机制至关重要。 需要更高的空间分辨率，例如囊泡运输、瞬时分子相互作用或 表观遗传变化的能力将衡量这些高度动态事件的微妙差异。 让我们的用户更好地掌握从健康系统到患病系统发生的分子变化。 多重模式的这种速度对于固定厚组织成像 (>200μm) 也至关重要，其中 大视场或厚切片可以在很短的时间内以超分辨率成像。 LSM980 带来了 SBP 目前不具备的额外成像优势。 LSM980 允许同时对全发射光谱（即所有荧光团）进行成像。 对弱样本更敏感，对背景噪声不太敏感，新添加的 Airyscan 联合反卷积处理提供 90nm 空间分辨率。 对于需要对厚样品进行超分辨率成像的项目至关重要，例如 细胞簇或果蝇胚胎 LSM980 将是圣地亚哥第一个可用的。 通过在 SBP 购买该仪器而实现的科学进步将 允许拥有 75% 可访问使用时间 (AUT) 的 12 个主要用户先前生成 无法获得的数据，这将进一步发展新的假设、新的资金以及两者 内部和外部合作。