JEOL JEM-1400 Tomographic Transmission EM with High-Resolution Digital Cameras

JEOL JEM-1400 配备高分辨率数码相机的断层扫描透射电子显微镜

• 批准号: 7838715
• 负责人: JOHN E RASH
• 金额: $ 80.43万
• 依托单位: COLORADO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-01 至 2012-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7838715
• 关键词: Actins; Archives; Binding; Boxing; Colorado; Electron Microscope; Electrons; Ensure; Exanthema; Excision; Film; Freeze Fracturing; Freezing; Frequencies; Funding; Gap Junctions; Image; Label; Laboratories; Liquid substance; Mechanics; Medicine; Membrane; Molecular; Monitor; National Center for Research Resources; Neurodegenerative Disorders; Neuronal Plasticity; Neurons; Nitrogen; Operative Surgical Procedures; Peer Review; Persons; Photography; Potassium Channel; Productivity; Publications; Publishing; Research Personnel; Resolution; Scanning; Schedule; Specimen; Staging; Surface; Surveys; Synapses; Time; Universities; Viral Proteins; Virion; Virus Receptors; Work; Wyoming; base; cofilin; college; digital; digital imaging; experience; neurotransmission; receptor; reconstruction; repaired; research study; retinal rods; stereoscopic; trafficking; transmission process

DESCRIPTION (provided by applicant): Five established and three new NIH-funded investigators from three departments at Colorado State University, plus one new investigator from the University of Wyoming, and one established investigator via a subcontract from Albert Einstein College of Medicine, request $804,260 in NCRR funds to purchase a JEOL JEM-1400 120 kV transmission electron microscope (TEM) equipped with a liquid nitrogen-cooled 700 tomographic specimen tilt holder, a fast-scan digital camera, and a high-resolution slow-scan digital camera. The new TEM will replace a 19-yr-old JEOL JEM-2000 EX-II 200 kV "workhorse" TEM that lacks a digital camera, has no tomographic capability, and has no automated stage drives and no automated photographic capability. Its "cut- film" camera has made >67,000 negatives (>67,000 mechanical cycles), requiring us to purchase three new camera boxes because the worn mechanical advance frequently jammed, irrevocably damaging three. The JEM-2000 has had increasing frequency of repairs ("down time" of 20-38 working days in each of the last five years. The requested bottom-mount high-resolution digital cameras, in combination with automated tilting and automated photography, are essential for tomographic analysis (up to 70 images for each reconstruction) and will greatly facilitate conventional photography. Digital images will be stored centrally for access online or offline without possibility of altering or deleting the archived originals. The video-rate 11MB camera will facilitate focusing and rapid tracking of image detail on the monitor during group analysis, and will be the primary camera for obtaining routine survey micrographs. Single-image capture of hundreds of virus particles using the 4K (16MB) high-resolution digital camera (as opposed to 2K or 1K cameras) will greatly reduce beam damage to virus/receptor-protein interactions in frozen thin films by reducing by 4x or 16x the number of exposures needed for tomographic analysis. On-line stereoscopic imaging, often at extreme tilts, is a necessity for interpreting surface topography and the "sidedness" of immunogold-labels in freeze-fracture replicas, rather than waiting until the next day for film to be developed. Removal of this delay between experiments will greatly accelerate work in several laboratories. The high productivity of current "major users" has resulted in >1100 published electron micrographs from the JEM-2000 in >50 peer-reviewed publications. TEM is essential for 6 NIH-funded major users investigating molecular composition of neuronal and glial membranes (Rash), ADF/cofilin-actin rods in neurodegenerative disease (Bamburg), synaptic and non-synaptic mechanisms of neuronal transmission (Reist; Rash), virus/protein molecular interactions (Hafenstein), regulators of K+ channel function and trafficking (Tamkun), and plasticity of neuronal gap junctions (Pereda/Rash). Based on 19 years of experience with the JEM-2000 TEM, a 10%-time administrative and technical person (Ms. Davidson) and a broadly-based Advisory Board will ensure continued efficient scheduling and operation of the upgraded replacement transmission electron microscope.

描述（由申请人提供）：来自科罗拉多州立大学三个系的五名新的NIH资助的调查员，以及一名来自怀俄明大学的新调查员，另一位通过来自阿尔伯特·爱因斯坦医学院的分包合同，一名成立的调查员，请求804,260美元的NCRR资金，以购买液体jem-1400 120 kem-1400 kem-1400 kem-1400 kem-1400 kem-1400 kem-1400 kem-1400 kvemose（ 700层析成像标本倾斜架，快速扫描的数码相机和高分辨率慢速扫描数码相机。新的TEM将取代缺乏数码相机，没有层析成绩的19岁JEOL JEL-2000 EX-II 200 kV“ Workhorse” TEM，并且没有自动化的舞台驱动器，也没有自动化的摄影功能。它的“切割胶片”相机造成了> 67,000个负面因素（> 67,000个机械周期），要求我们购买三个新的相机盒，因为磨损的机械进步经常被困住，不可避免地损坏了三个。 JEM-200000的维修频率越来越高（过去五年中的每个工作日的“停机时间”为20-38个工作日。所需的底部安装高分辨率的数码相机，结合自动化和自动摄影的结合，对于层析成像分析至关重要（对于每次重建而言，最多可访问70张图像），并且将访问数字图像。删除归档的原始镜头。通过减少4倍或16倍的层次，在线倾斜度的直接成像是在极端倾斜的情况下进行的，是解释表面地形的必要性，而在冻结式复制中的“侧面”是造成的。 current "major users" has resulted in >1100 published electron micrographs from the JEM-2000 in >50 peer-reviewed publications. TEM is essential for 6 NIH-funded major users investigating molecular composition of neuronal and glial membranes (Rash), ADF/cofilin-actin rods in neurodegenerative disease (Bamburg), synaptic and non-synaptic mechanisms of neuronal transmission （REIST;皮疹），病毒/蛋白质分子相互作用（Hafenstein），K+通道功能和运输的调节剂（Tamkun）以及神经元间连接的可塑性以及基于19年的JEM-2000 TEM的经验，并确保了10％的行政和技术委员（MS. Ms. David S. Ms. Ms. Ms.-Ms.-Ms.升级的替换透射电子显微镜。