Cryo Upgrade of UMB EM Core Facility

UMB EM 核心设施的冷冻升级

• 批准号: 7794116
• 负责人: RU-CHING HSIA
• 金额: $ 42.07万
• 依托单位: UNIVERSITY OF MARYLAND BALTIMORE
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-08-01 至 2012-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7794116
• 关键词: Animals; Antigens; Baltimore; Biological; Biological Preservation; Biotechnology; Cells; Communities; Complement; Core Facility; Correlative Study; County; Cryoelectron Microscopy; Detection; Devices; Dose; Electron Microscope; Electron Microscopy; Equipment; Freeze Substitution; Freezing; Funding; Grant; Heavy Metals; Ice; Imaging technology; Immunogold Technics; Institutes; Lasers; Life; Light; Maryland; Methods; Microscope; Plant Resins; Preparation; Research; Research Institute; Research Personnel; Research Project Grants; Resolution; Sampling; Scanning; Specimen; Staining method; Stains; System; Techniques; Tissues; Universities; base; cellular imaging; cold temperature; instrument; microorganism; pressure; protein complex; transmission process

DESCRIPTION (provided by applicant): We request funds toward the purchase of cryo-EM sample preparation equipment, including a high pressure freezer (HPF), an automatic freeze substitution system, a cryo-ultramicrotome, a plunge freezing device and the cryo upgrade of an existing transmission electron microscope (TEM) Tecnai T12 (FEI). These instruments will be integrated into a newly re-organized electron microscopy (EM) core facility, the only core facility for researchers at the University of Maryland Baltimore (UMB), University of Maryland Biotechnology Institute (UMBI), University of Maryland Baltimore County (UMBC) and other neighboring research institutes. It is well established that cryo sample preparation techniques offers better antigen preservation and enhanced spatial resolution of ultrastructures. The cryo TEM upgrade, incorporating a cryo sample holder and low dose imaging technologies will allow researchers to observe frozen hydrated biological samples at high resolution, an option that has not been available to researchers at UMB and other surrounding institutes. Many of the major user research projects (Drs. Bavoil, Nataro, Donnenberg, Russell and Bloch) described in this application will utilize immunogold techniques following HPF, freeze substitution and resin embedding at low temperature to enhance antigen detection and ultrastructural preservation. Major users Drs. Donnenberg and Bavoil will use the plunge freeze device to prepare purified protein complexes or microorganisms in vitreous ice and subsequently perform high resolution ultrastructure analysis in the cryo TEM without the use of heavy metal staining. Furthermore, the rapid transfer system used in combination with the HPF allows correlative studies of the same specimen by both light and electron microscopy. This feature complements the capability of live cell imaging of the newly acquired Zeiss Duo laser scanning confocal microscope (purchased in 2008 via SIG 1S10RR02454801, Thomas Blanpied, investigator, major user on this application) and the growing number of research projects studying live cells on the UMB/UMBI campus. Major users Drs. Blanpied and Martin plan to use this feature to enhance their live cell-imaging research. This grant is based upon funded projects for which the cryo sample preparation and cryo EM techniques offer substantial improvements over conventional EM methods for animal cells, whole tissues and microorganisms. Other ongoing projects in this research community will also immediately benefit from the expanded capability of the EM core facility.

描述（由申请人提供）：我们请求资金用于购买冷冻电镜样品制备设备，包括高压冷冻机 (HPF)、自动冷冻替代系统、冷冻超薄切片机、插入式冷冻装置和冷冻升级现有的透射电子显微镜 (TEM) Tecnai T12 (FEI)。这些仪器将集成到新重组的电子显微镜 (EM) 核心设施中，这是马里兰大学巴尔的摩分校 (UMB)、马里兰大学生物技术研究所 (UMBI)、马里兰大学巴尔的摩县分校 (UMB) 的研究人员的唯一核心设施。 UMBC）和其他邻近的研究机构。众所周知，冷冻样品制备技术可以更好地保存抗原并增强超微结构的空间分辨率。冷冻 TEM 升级结合了冷冻样品支架和低剂量成像技术，将使研究人员能够以高分辨率观察冷冻水合生物样品，这是 UMB 和其他周边研究所的研究人员无法获得的选项。本申请中描述的许多主要用户研究项目（Bavoil、Nataro、Donnenberg、Russell 和 Bloch）将利用 HPF、冷冻替代和低温树脂包埋后的免疫金技术来增强抗原检测和超微结构保存。主要用户 Drs. Donnenberg 和 Bavoil 将使用切入式冷冻装置在玻璃冰中制备纯化的蛋白质复合物或微生物，然后在冷冻 TEM 中进行高分辨率超微结构分析，而无需使用重金属染色。此外，快速转移系统与 HPF 结合使用，可以通过光学显微镜和电子显微镜对同一样本进行相关研究。这一功能补充了新购买的 Zeiss Duo 激光扫描共聚焦显微镜（2008 年通过 SIG 1S10RR02454801 购买，Thomas Blanpied，研究员，该应用的主要用户）的活细胞成像功能，以及越来越多的研究活细胞的研究项目UMB/UMBI 校园。主要用户 Drs. Blanpied 和 Martin 计划利用这一功能来加强他们的活细胞成像研究。该资助基于所资助的项目，这些项目的冷冻样品制备和冷冻电镜技术相对于动物细胞、整个组织和微生物的传统电镜方法提供了实质性改进。该研究界正在进行的其他项目也将立即受益于 EM 核心设施的扩展功能。

项目成果

Defective sarcomere assembly in smyd1a and smyd1b zebrafish mutants.

smyd1a 和 smyd1b 斑马鱼突变体中肌节组装缺陷。

• 发表时间: 2019-05
• 作者: Cai, Mengxin;Han, Lichen;Liu, Lusha;He, Feng;Chu, Wuying;Zhang, Jianshe;Tian, Zhenjun;Du, Shaojun
• 通讯作者: Du, Shaojun

SIRT3, a metabolic target linked to ataxia-telangiectasia mutated (ATM) gene deficiency in diffuse large B-cell lymphoma.

SIRT3，一种与弥漫性大 B 细胞淋巴瘤中共济失调毛细血管扩张突变 (ATM) 基因缺陷相关的代谢靶点。

• 发表时间: 2020
• 影响因子: 4.6
• 作者: Bhalla, Kavita;Jaber, Sausan;Reagan, Kayla;Hamburg, Arielle;Underwood, Karen F;Jhajharia, Aditya;Singh, Maninder;Bhandary, Binny;Bhat, Shambhu;Nanaji, Nahid M;Hisa, Ruching;McCracken, Carrie;Creasy, Heather Huot;Lapidus, Rena G;Kingsbury, Ta
• 通讯作者: Kingsbury, Ta

Separation, Sizing, and Quantitation of Engineered Nanoparticles in an Organism Model Using Inductively Coupled Plasma Mass Spectrometry and Image Analysis.

使用电感耦合等离子体质谱和图像分析对生物体模型中的工程纳米颗粒进行分离、分级和定量。

• 发表时间: 2017
• 影响因子: 17.1
• 作者: Johnson, Monique E;Hanna, Shannon K;Montoro Bustos, Antonio R;Sims, Christopher M;Elliott, Lindsay C C;Lingayat, Akshay;Johnston, Adrian C;Nikoobakht, Babak;Elliott, John T;Holbrook, R David;Scott, Keana C K;Murphy, Karen E;Petersen, Elijah J
• 通讯作者: Petersen, Elijah J