Stanford-SLAC CryoET Specimen Preparation Service Center (SCSC)

斯坦福-SLAC CryoET 标本制备服务中心 (SCSC)

• 批准号: 10818212
• 负责人: Wah Chiu
• 金额: $ 330.19万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-16 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10818212
• 关键词: Academia; Adopted; Advisory Committees; Area; Award; Biocompatible Materials; Biological; Cells; Collaborations; Communication; Communities; Complex; Cryo-electron tomography; Cryoelectron Microscopy; Data Collection; Dedications; Educational Curriculum; Educational workshop; Electron Microscope; Equipment; Fluorescence Microscopy; Freezing; Funding; Goals; Hydration status; Ice; In Situ; Industry; Infrastructure; Institution; Ions; Laboratories; Lifting; Macromolecular Complexes; Methods; Microscope; Organelles; Preparation; Process; Professional Organizations; Protocols documentation; Recurrence; Request for Applications; Research; Resource Allocation; Resources; Sampling; Scanning; Scanning Electron Microscopy; Services; Specimen; Subcellular structure; Techniques; Technology; Thick; Thinness; Tissue Sample; Tissues; Titan; Training; Training Activity; United States National Institutes of Health; Work; cryogenics; electron tomography; experimental study; imaging modality; innovative technologies; nanometer resolution; response; structural biology; symposium; tool; web portal

Project Summary In response to the NIH RFA-RM-19-009, we propose to establish the Stanford-SLAC CryoET Specimen Preparation Service Center (SCSC) to accomplish four specific aims: (1) establish a platform, which is available to the scientific community at large, to streamline the preparation of samples suitable for downstream cryogenic electron tomography (cryoET) data collection; (2) provide access to advanced cryo-specimen preparation techniques for a wide range of samples: macromolecular complexes, microcrystals of biological materials, cell lysates, organelles, cells and tissues; (3) generate a training curriculum for new users on the preparation of frozen, hydrated biological specimens, with the option of carrying out correlative cryo-fluorescence light microscopy (cryoFLM), cryo-focused ion beam scanning electron microscopy (cryoFIB), and cryoET; and (4) adopt new methods for sample preparation using innovative technologies developed elsewhere. In this proposal, we will leverage our existing facilities and in-house expertise in each of the following imaging modalities and their integration into a correlated sample preparation workflow: cryoFLM, cryoFIB, and cryoET. We will establish all the necessary equipment dedicated to the proposed service and training activities. We will prepare video curriculum materials to train new users in the aforementioned protocols. We will adapt our existing infrastructure to manage project administration and resource allocations. Administrative support is in place to help users with lodging arrangements, as well as laboratory on-boarding and specimen biosafety approvals. We have a well set- up communication infrastructure for remote users to participate in actual experiments, particularly for cryoFLM, cryoFIB, and cryoET. We will provide access to existing Titan Krios microscopes to evaluate whether the prepared samples are ready for data collection in the associated Hub, which will be set up at another institution with separate NIH support. We will make our existing Stanford- SLAC cryo-specimen preparation equipment available on a limited basis immediately after this proposal is funded. We anticipate serving ~40 users per year once the Center is fully operational in early Year 2 of the award period. We will also offer regular hands- on workshops to train ~10-12 new users per year. We will disseminate our resources to the broad community via a web portal and booths at professional societies’ annual conferences. The number of users will increase in subsequent years as we and our recurring users optimize the use of the Center’s resources. In Years 3-6, we will work closely with collaborators pioneering advanced protocols such as semi-automated and automated cryoFIB and “lift-out” for tissue samples to implement them in the proposed Center. Lastly, we will assemble a Scientific Advisory Committee of experts to guide the practices of our Service Center and the ongoing implementation of cutting- edge technologies.

项目总结