A Multi-user Super Resolution Microscope for Developmental Biology

用于发育生物学的多用户超分辨率显微镜

基本信息

  • 批准号:
    7836356
  • 负责人:
  • 金额:
    $ 104.95万
  • 依托单位:
  • 依托单位国家:
    美国
  • 项目类别:
  • 财政年份:
    2010
  • 资助国家:
    美国
  • 起止时间:
    2010-09-11 至 2012-09-10
  • 项目状态:
    已结题

项目摘要

DESCRIPTION (provided by applicant): Understanding the molecular basis of developmental mechanisms ultimately requires precise knowledge of events in space and time. While the clearest view of subcellular organization has come from electron microscopy, its utility in understanding mechanism is burdened by the difficulty of mapping molecule to structure. Recent advances now offer us nanometer-scale resolution through technologies like STED, PALM/STORM and SIM that bring the resolution of light microscopy below the Abbe limit. This proposal requests support for the purchase of a Zeiss LSM710 PAL-M/SIM microscope. This instrument offers three imaging modalities; laser scanning confocal microscopy, Photo Activated Localization Microscopy (PAL-M), and Structured Illumination Microscopy (SIM). With this combination, one can obtain in a single specimen a broad 3D perspective by confocal microscopy and, in selected regions, sub-diffraction limit resolution with SIM and PAL-M. In combination with a wide array of multi-color fluorescent molecular probes, these are powerful technologies that open vast new areas of inquiry. In this proposal, the Principal Investigator Sam Kunes and five group leaders describe developmental biology projects that implement this technology. Developmental processes are a particularly suitable subject for super-resolution microscopy since they are governed and fine-tuned by spatial cues, many of which act in the sub-diffraction limited range. Three groups, Kunes, Lichtman and Dulac, propose to study the development of the ultrastructure of synaptic connections. Kunes will investigate how a developmental pathway is re-deployed to control long-term memory control axonal and dendritic branching, synapse number, and synapse morphology in the mature fruitfly brain. Lichtman plans to decipher the development of complex patterns of axonal circuitry and synaptic connectivity at the neuromuscular junction, and between neurons in the parasympathetic ganglia and cerebral cortex, using Brainbow mice. The Dulac laboratory will investigate the role of the Major Histocompatability M10 family of proteins in the developmental plasticity of vomeronasal organ sensory neuron axon terminals. Two groups, McMahon and Schier, propose to visualize morphogen movement at high resolution; McMahon will focus on Sonic Hedgehog, as it patterns the ventral neural tube during CNS development, while Schier proposes to image the morphogen Squint and its co-localization with other components of the Nodal signaling pathway during Zebrafish development. The Mango laboratory seeks to understand how the packaging of chromatin is modulated during development, in order to understand the transition from developmental plasticity to cell fate commitment. The instrument will be incorporated into a well-established imaging facility, where it will receive full-time technical support and oversight, support for maintenance and managed usage, thus providing long term availability to the co-PI group and the broader community of microscopists.
描述(由申请人提供):理解发育机制的分子基础最终需要对空间和时间事件的精确了解。虽然亚细胞组织的最清晰视图来自电子显微镜,但其在理解机制方面的效用却受到将分子映射到结构的困难的影响。最近的进展现在通过 STED、PALM/STORM 和 SIM 等技术为我们提供了纳米级分辨率,使光学显微镜的分辨率低于阿贝极限。该提案请求支持购买 Zeiss LSM710 PAL-M/SIM 显微镜。该仪器提供三种成像模式;激光扫描共焦显微镜、光激活定位显微镜 (PAL-M) 和结构照明显微镜 (SIM)。通过这种组合,人们可以通过共焦显微镜在单个样本中获得广泛的 3D 视角,并在选定区域中使用 SIM 和 PAL-M 获得亚衍射极限分辨率。与各种多色荧光分子探针相结合,这些强大的技术开辟了广阔的新研究领域。在这份提案中,首席研究员 Sam Kunes 和五名小组负责人描述了实施该技术的发育生物学项目。发育过程是超分辨率显微镜特别适合的主题,因为它们受到空间线索的控制和微调,其中许多空间线索在亚衍射有限范围内起作用。 Kunes、Lichtman 和 Dulac 三个小组提议研究突触连接超微结构的发展。 Kunes 将研究如何重新部署发育途径来控制成熟果蝇大脑中的长期记忆控制轴突和树突分支、突触数量和突触形态。 Lichtman 计划使用 Brainbow 小鼠破译神经肌肉接头处以及副交感神经节和大脑皮层神经元之间轴突回路和突触连接的复杂模式的发展。 Dulac 实验室将研究主要组织相容性 M10 蛋白家族在犁鼻器感觉神经元轴突末端发育可塑性中的作用。麦克马洪和席尔这两个小组提出以高分辨率可视化形态发生素的运动。 McMahon 将重点关注 Sonic Hedgehog,因为它在 CNS 发育过程中模拟腹侧神经管,而 Schier 则建议对斑马鱼发育过程中的形态素 Squint 及其与 Nodal 信号通路其他组件的共定位进行成像。 Mango 实验室致力于了解发育过程中染色质包装的调节方式,以了解从发育可塑性到细胞命运决定的转变。该仪器将被整合到一个完善的成像设施中,在那里它将获得全职技术支持和监督、维护和管理使用支持,从而为联合 PI 小组和更广泛的显微镜专家社区提供长期可用性。

项目成果

期刊论文数量(2)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
Transit and integration of extracellular mitochondria in human heart cells.
  • DOI:
    10.1038/s41598-017-17813-0
  • 发表时间:
    2017-12-12
  • 期刊:
  • 影响因子:
    4.6
  • 作者:
    Cowan DB;Yao R;Thedsanamoorthy JK;Zurakowski D;Del Nido PJ;McCully JD
  • 通讯作者:
    McCully JD
Targeting host mitochondria: A role for the Trypanosoma cruzi amastigote flagellum.
  • DOI:
    10.1111/cmi.12807
  • 发表时间:
    2018-03
  • 期刊:
  • 影响因子:
    3.4
  • 作者:
    Lentini G;Dos Santos Pacheco N;Burleigh BA
  • 通讯作者:
    Burleigh BA
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Samuel M Kunes其他文献

Samuel M Kunes的其他文献

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{{ truncateString('Samuel M Kunes', 18)}}的其他基金

Mechanisms of Morphogen Secretion in Visual System Development and Disease
视觉系统发育和疾病中形态原分泌的机制
  • 批准号:
    8621495
  • 财政年份:
    2014
  • 资助金额:
    $ 104.95万
  • 项目类别:
Mechanisms of Morphogen Secretion in Visual System Development and Disease
视觉系统发育和疾病中形态原分泌的机制
  • 批准号:
    8788031
  • 财政年份:
    2014
  • 资助金额:
    $ 104.95万
  • 项目类别:
Mechanisms of Morphogen Secretion in Visual System Development and Disease
视觉系统发育和疾病中形态原分泌的机制
  • 批准号:
    9195099
  • 财政年份:
    2014
  • 资助金额:
    $ 104.95万
  • 项目类别:
Mechanisms of Morphogen Secretion in Visual System Development and Disease
视觉系统发育和疾病中形态原分泌的机制
  • 批准号:
    8987570
  • 财政年份:
    2014
  • 资助金额:
    $ 104.95万
  • 项目类别:
Regulation of Memory by the microRNA/RISC Pathway
microRNA/RISC 通路对记忆的调节
  • 批准号:
    7583125
  • 财政年份:
    2009
  • 资助金额:
    $ 104.95万
  • 项目类别:
Regulation of Memory by the microRNA/RISC Pathway
microRNA/RISC 通路对记忆的调节
  • 批准号:
    7753917
  • 财政年份:
    2009
  • 资助金额:
    $ 104.95万
  • 项目类别:
Regulation of Memory by the microRNA/RISC Pathway
microRNA/RISC 通路对记忆的调节
  • 批准号:
    8197612
  • 财政年份:
    2009
  • 资助金额:
    $ 104.95万
  • 项目类别:
Regulation of Memory by the microRNA/RISC Pathway
microRNA/RISC 通路对记忆的调节
  • 批准号:
    7995161
  • 财政年份:
    2009
  • 资助金额:
    $ 104.95万
  • 项目类别:
Regulation of Memory by the microRNA/RISC Pathway
microRNA/RISC 通路对记忆的调节
  • 批准号:
    8391720
  • 财政年份:
    2009
  • 资助金额:
    $ 104.95万
  • 项目类别:
Synaptic Structural Plasticity in the Drosophila Brain
果蝇大脑的突触结构可塑性
  • 批准号:
    7146182
  • 财政年份:
    2006
  • 资助金额:
    $ 104.95万
  • 项目类别:

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