Novel TIRF microscopy analyzing trafficking & signaling at the cell cortex

新型 TIRF 显微镜分析贩运

• 批准号: 7892704
• 负责人: Derek K. Toomre
• 金额: $ 5.05万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-08-03 至 2010-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7892704
• 关键词: 1-Phosphatidylinositol 3-Kinase; Abbreviations; Accounting; Acoustics; Address; Adipocytes; Affect; Algorithms; Area; Arts; Attenuated; Automobile Driving; Award; Back; Binding; Biochemical; Biochemistry; Biological; Biology; Boxing; Buffers; Caliber; Calibration; Cell Line; Cell membrane; Cell surface; Cells; Cellular biology; Clathrin; Cluster Analysis; Collaborations; Collection; Collimator; Color; Coma; Communities; Complex; Computational Biology; Computer Vision Systems; Computers; Conflict (Psychology); Confocal Microscopy; Coupled; Coupling; Cues; Cytoskeleton; Data; Data Set; Defect; Development; Diabetes Mellitus; Diffusion; Dimensions; Dimerization; Disadvantaged; Discipline; Docking; Down-Regulation; Drops; Dyes; Employee Strikes; Endocytosis; Engineering; Ensure; Environment; Event; Exocytosis; Eye; Face; Feedback; Fiber; Figs - dietary; Flare; Fluorescein-5-isothiocyanate; Fluorescence; Fluorescence Microscopy; Fluorescence Recovery After Photobleaching; Fluorescent Dyes; Functional disorder; Funding; Genetic Screening; Germany; Glass; Glucose Transporter; Glycerol; Goals; Grant; Image; Image Analysis; Imagery; Incidence; Insulin; Interdisciplinary Study; Investments; Joints; Kinetics; Knowledge; Label; Laboratories; Lasers; Learning; Legal patent; Length; Life; Light; Lighting; Link; Lipids; Location; Macromolecular Complexes; Malignant Neoplasms; Maps; Masks; Measures; Mediating; Membrane; Membrane Microdomains; Membrane Protein Traffic; Methodology; Methods; Microscope; Microscopy; Microtubules; Modeling; Molecular; Monitor; Morphologic artifacts; Motivation; Motor; Nature; Non-Insulin-Dependent Diabetes Mellitus; Optics; Organelles; PTEN gene; Paper; Parasites; Pathway interactions; Penetration; Performance; Phosphatidylinositols; Phosphotransferases; Photobleaching; Physiologic pulse; Planet Mars; Play; Positioning Attribute; Postdoctoral Fellow; Private Sector; Probability; Process; Proteins; Publications; Pupil; Quantum Dots; RNA Interference; Radial; Randomized; Reagent; Recruitment Activity; Refractive Indices; Regulation; Relative (related person); Reporter; Research; Research Infrastructure; Research Personnel; Research Support; Resolution; Risk; Role; Sampling; Scanning; Science; Scientist; Seminal; Series; Side; Signal Pathway; Signal Transduction; Silicon Dioxide; Simulate; Site; Small Interfering RNA; Solid; Solutions; Sorting - Cell Movement; Source; Spain; Spatial Distribution; Specific qualifier value; Specimen; Speed; Spottings; Structure; Surface; System; Techniques; Technology; Testing; Text; Thick; Time; Total Internal Reflection Fluorescent; Touch sensation; Training; Transfection; Tubulin; Vesicle; Visual; Wolves; Work; analytical method; basal insulin; base; blood glucose regulation; cell cortex; cell motility; cell type; cellular imaging; density; design; extracellular; flexibility; flotillin; fluorescence imaging; fluorescence microscope; fluorophore; handbook; holistic approach; image processing; improved; innovation; insight; instrument; instrumentation; insulin signaling; interest; lens; medical schools; meetings; micromanipulator; migration; millisecond; nanometer; novel; object shape; photoactivation; prototype; receptor; research study; response; scaffold; simulation; single molecule; success; tool; trafficking; trans-Golgi Network; trend; user-friendly; virtual

In 3-month paid research experiences, over two summers, 4 talented undergraduate engineering and computer science students will be recruited to contribute in a substantial way to the progress of sub-aims of my parent New Innovator (DP2) Award; the DP2's main specific aims are to develop novel microscopes (Aim 1) and analytical tools (Aim 2) to study membrane trafficking at the cell cortex (Aim 3). Specifically, students will work on one of two projects: i) Design and construct and improve electronic circuits to control the galvometric XY mirror on our multi-angle FRAP/TIRFM (Aim 1B) and/or new auto-sensing calibration devices (Aim 2) or ii) Develop models or software to visualize and track vesicles in 3D by multi-angle TIRFM (Aim 2 ). Their advances will directly benefits goals of the DP2 and the new electronics and software's performance will be benchmarked and iteratively improved. Training and oversight will come from the PI and the DP2-supported senior scientist, Dr. Polejaev. Students will benefit from the infrastructure of the Yale 'CINEMA' lab imaging center, which is under the PI's directorship. Two outstanding US engineering students have already been identified, Noah Pestana and Isaac Anderson, both of whom have expressed a high interest in participating this summer on projects (i) and (ii), respectively. In the second summer, a particular emphasis will be made to recruit minority undergraduate students through the Yale 'STARS' program for minorities. All proposed activity are within the scope of the parent DP2 and capitalizes on early successes and will accelerate the tempo of two of the approved specific aims. Consistent with the recovery act's goal, this funding will provide full-time summer employment for 4 undergraduate students and accelerate scientific achievement of the parent DP2 award.

在为期3个月的有偿研究经验中，在两个夏天里，将招募4名有才华的工程和计算机科学本科生，为我的父母新创新者（DP 2）奖的子目标的进展做出实质性贡献; DP 2的主要具体目标是开发新型显微镜（目标1）和分析工具（目标2），以研究细胞皮层的膜运输（目标3）。 具体来说，学生将从事两个项目之一：i）设计，构建和改进电子电路，以控制我们的多角度FRAP/TIRFM（目标1B）和/或新的自动传感校准设备（目标2）或ii）开发模型或软件，通过多角度TIRFM（目标2）在3D中可视化和跟踪囊泡。他们的进步将直接有利于DP 2的目标，新的电子和软件的性能将被基准测试和迭代改进。培训和监督将来自PI和DP 2支持的高级科学家Polejaev博士。 学生将受益于耶鲁大学'CINEMA'实验室成像中心的基础设施，该中心由PI领导。 已经确定了两名优秀的美国工程专业学生，Noah Pestana和Isaac安德森，他们都表示有兴趣参加今年夏天的项目（i）和（ii）。在第二个夏天，将特别强调通过耶鲁大学的少数民族“明星”计划招收少数民族本科生。 所有拟议的活动都在母项目发展计划2的范围内，并利用早期的成功，将加快两个核定具体目标的克里思。与复苏法案的目标一致，这笔资金将为4名本科生提供全职夏季就业机会，并加速父母DP 2奖的科学成就。

项目成果

Live-cell imaging of exocyst links its spatiotemporal dynamics to various stages of vesicle fusion.

• DOI: 10.1083/jcb.201212103
• 发表时间: 2013-05-27
• 期刊: The Journal of cell biology
• 作者: Rivera-Molina F;Toomre D
• 通讯作者: Toomre D