ShEEP Request for Nikon TIRF STORM microscope

SheEEP 请求尼康 TIRF STORM 显微镜

• 批准号: 9795504
• 负责人: STEPHEN E MCGOWAN
• 金额: --
• 依托单位: IOWA CITY VA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-01-01 至 2019-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9795504
• 关键词: Adaptor Signaling Protein; Adhesions; Algorithms; Automation; Bacteria; Budgets; Carbon Dioxide; Cell Communication; Cell Differentiation process; Cell Surface Receptors; Cell membrane; Cell surface; Cells; Chromatin; Cities; Communicable Diseases; Computer software; Cytomegalovirus; Cytoskeleton; Defect; Detection; Development; Diabetes Mellitus; Disease; Disease Progression; Electrons; Elements; Endosomes; Endothelium; Environment; Equilibrium; Exhibits; Extracellular Matrix; Eye; Fluorescence; Funding; Future; Gene Expression; Genomics; Glass; Glaucoma; HIV; Health; Heart; Hepatitis C; Hepatitis C virus; Host Defense; Humidity; Image; Immersion Investigative Technique; Incubators; Individual; Institution; Insulin; Investigation; Iowa; Knowledge; Lasers; Learning; Legionella; Leishmaniasis; Lighting; Liver; Lung; Lung diseases; Malignant Neoplasms; Mechanics; Medical center; Microbiology; Microprocessor; Microscope; Microscopy; Microvascular Dysfunction; Military Personnel; Morphology; Motor; Movement; Noise; Obesity; Oils; Ophthalmology; Optics; Organ; Organelles; Parasites; Perfusion; Pericytes; Peripheral; Phenotype; Phosphorylation; Photons; Physiology; Population; Process; Proteins; Proteomics; Pulmonary Emphysema; Reagent; Receptor Signaling; Recycling; Research; Research Design; Research Personnel; Resolution; Sheep; Signal Transduction; Skin; Speed; Structure; System; T-Cell Receptor; Techniques; Technology; Temperature; Time; Tissues; Universities; Vendor; Veterans; Viral; Viral Proteins; Virus; Wound Healing; adiponectin; antigen processing; cell motility; chronic infection; density; fluorescence imaging; image reconstruction; image registration; in vivo; instrument; intravital microscopy; knowledge base; lens; light intensity; live cell imaging; microscopic imaging; novel; optical emission; pathogen; programs; protein complex; quantum; receptor; reconstruction; serial imaging; single molecule; solid state; temporal measurement; trafficking

Super-resolution microscopy has opened unprecedented opportunities to image single molecules or groups of molecules with resolution below 50 nm. However microscopes with these capabilities, if purchased from a commercial vendor as ready to assemble, are beyond the budgets of most individual investigators. And cutting expenses by building one's own requires an understanding of optics which is beyond the knowledge of most biologists. Multiple investigators associated with the Iowa City VA Medical Center have reached a point in their investigations where applying super-resolution microscopy to living cells is required to move their research forward. This research is targeted towards diseases which are relevant to US military veterans: pulmonary emphysema, alterations in host defense observed in chronic infections such as hepatitis C, cytomegalovirus, or Leishmaniasis, glaucoma and obesity. There is currently no microscope on the VA or University of Iowa campuses which will enable these types of studies. Therefore, we are requesting a Nikon Eclipse Ti-e2 inverted microscope equipped with total internal reflection (TIRF) and stochastic optical reconstruction microscopy (STORM) modules. TIRF optics will benefit those studying receptors which signal by assembling adapter proteins at the plasma membrane (T-cell receptor, Drs. Bishop and Stapleton) or retain active signaling after entering endosomes (Dr. McGowan). Combining TIRF and STORM will enable Drs. McGowan, Allen and Anderson to study the assembly of podosomes or remodeling of the sub-cortical cytoskeleton. STORM will enable Drs. Wilson, Meier, and Stapleton to observe the assembly of proteins from pathogens with those from host cells to learn how viruses or parasites evade host defenses. Dr. Meier will use STORM to study targeted degradation of viral or mammalian proteins or the interaction of viral proteins with chromatin. All of these projects require multicolor fluorescent imaging, at high resolution, to detect and localize photons, at rapid frame rates and with high directional and temporal precision. We have chosen the Nikon H-TIRF/N- STORM system because it meets all of these requirements in a package which provides these features in a largely automated format, which is highly desirable for a multi-user instrument. To enable live cell imaging the microscope will be equipped with a stage-top incubator to control the environment and allow reagents to be introduce without altering the field of view. We are also requesting powerful PC workstations and Nikon software which is required to acquire and analyze images. A more complete description of the instrument may be found in the Specific Aims section. Because this package involves emerging state-of the art technology, we hope that it will also enhance the research programs of new investigators as they join the VA research program and pursue novel projects tailored to the future health needs of US veterans.

超分辨率显微镜已经为成像单分子或组的前所未有的机会开放 分辨率低于50 nm的分子。但是具有这些功能的显微镜，如果从 准备组装的商业供应商超出了大多数个人调查人员的预算。和切割 通过自己构建的费用需要对光学的理解，这超出了大多数知识 生物学家。与爱荷华城弗吉尼亚州医疗中心相关的多个调查人员已经达到了他们的点 需要对活细胞应用超分辨率显微镜的调查才能移动他们的研究 向前。这项研究针对与美国退伍军人相关的疾病：肺 肺气肿，在慢性感染中观察到的宿主防御改变，例如丙型肝炎，巨细胞病毒或 利什曼病，青光眼和肥胖症。爱荷华大学目前没有显微镜 可以实现这类研究的校园。因此，我们要求Nikon Eclipse Ti-e2 配备总内反射（TIRF）和随机光学重建的倒置显微镜 显微镜（风暴）模块。 TIRF光学元件将使那些通过组装来发出信号的研究受体受益 质膜（T细胞受体，主教和Stapleton博士）处的衔接蛋白或保持活性 进入内体后的信号（McGowan博士）。结合TIRF和Storm将使Drs启用。麦高恩， 艾伦（Allen）和安德森（Anderson）研究了足体的组装或皮下细胞骨架的重塑。 风暴将使Drs启用。威尔逊，梅尔和斯塔普尔顿观察病原体的蛋白质组装 从宿主细胞中学习病毒或寄生虫如何逃避宿主防御的人。 Meier博士将使用风暴 研究靶向病毒或哺乳动物蛋白的降解或病毒蛋白与染色质的相互作用。全部 在这些项目中，需要高分辨率的多色荧光成像来检测和定位光子 快速帧速率以及具有高方向和时间精度。我们选择了尼康H-Tirf/n- 风暴系统是因为它在包装中满足所有这些要求，该包装提供了这些功能 在很大程度上是自动化的格式，这对于多用户仪器非常需要。启用活细胞成像 显微镜将配备一个台式孵化器，以控制环境并允许试剂为 介绍而无需更改视野。我们还要求强大的PC工作站和尼康 获取和分析图像所需的软件。对乐器的更完整描述可能 在特定目标部分中找到。因为此包涉及新兴的最新技术，所以我们 希望在新调查人员加入VA研究计划时，它还可以增强他们的研究计划 并从事满足美国退伍军人未来健康需求的新颖项目。