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

项目摘要 Sanford Burnham Prebys（SBP）拥有一系列非常广泛的实验室，包括 癌症、衰老和与年龄相关的疾病，如神经变性、细胞和组织稳态， 功能、代谢和干细胞生物学。我们真的相信， 感兴趣的是，使用染料或荧光团，在它们的空间和时间位置的上下文中， 细胞或组织是理解正常与患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。然而，在这方面， 空间分辨能力的增加通常需要牺牲时间分辨率 可以观察到荧光团的分子动力学的速度，这需要仔细 选择满足实验需要所需的超分辨率模式。为此目的， SBP请求支持带有Airyscan 2的蔡司LSM 980。LSM980能够 同时成像多达4个不同的荧光团， 发射光谱的空间分辨率增加了两倍。仅限多路复用模式 LSM 980上提供，可将图像采集速度提高25倍， 典型的点扫描共焦显微镜的采集速率，仍然提供了近两倍的 提高空间分辨率。这种速度对于研究高度动态的机制至关重要， 需要更高的空间分辨率，如囊泡运输，瞬时分子相互作用或 表观遗传变化在这些高度动态的事件中测量细微差异的能力将 让我们的用户更好地掌握从健康到患病系统发生的分子变化。 多路模式的这种速度对于固定厚组织成像（>200µm）也至关重要， 大视场或厚切片可以在一小部分时间内以超分辨率成像。的 LSM 980带来了SBP目前无法提供的额外成像优势。类星体探测器 允许对全部发射光谱即所有荧光团同时成像。LSM980是 对弱样本更敏感，对背景噪声不太敏感， Airyscan联合反卷积处理提供90nm的空间分辨率。这些特性使得 对于需要对厚样品，例如类器官， 细胞簇或果蝇胚胎。由于LSM 980将是第一款可用于圣地亚哥的 在科学界，通过在SBP获得这一仪器所取得的科学进步将 允许拥有75%可访问使用时间（AUT）的12个主要用户生成先前 无法获得的数据，这将进一步发展新的假设，新的资金， 内部和外部合作。

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.