Ultra-Precise Laser Surgery to study Cell Biomechanics

超精密激光手术研究细胞生物力学

• 批准号: 7282382
• 负责人: ALAN J HUNT
• 金额: $ 24.91万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-09-01 至 2009-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7282382
• 关键词: Ablation; Achievement; Address; Antimitotic Agents; Architecture; Binding; Biological; Biology; Biomechanics; Cell Cycle; Cell division; Cells; Cellular Structures; Cellular biology; Centrioles; Centrosome; Characteristics; Chemistry; Chromosomes; Chromosomes, Human, 1-3; Communities; Cytoskeleton; Development; Disruption; Dose; Facility Construction Funding Category; Failure; Future; Gene Proteins; Genes; Genetic; Genetic Materials; Genome; Goals; Growth Cones; Heating; Indium; Kinetochores; Knock-out; Laser Surgery; Lasers; Learning; Light; Location; Measurement; Mechanics; Methods; Microfluidics; Microsurgery; Microtubules; Mitosis; Mitotic; Mitotic Chromosome; Molecular Genetics; Movement; Neurites; Optics; Organism; Paclitaxel; Pharmaceutical Preparations; Phenotype; Physiologic pulse; Process; Property; Pulse taking; Range; Rate; Research; Role; Science; Structure; Structure-Activity Relationship; Subcellular structure; Techniques; Technology; Testing; Therapeutic; absorption; base; cell motility; chemotherapy; chromosome movement; concept; design; improved; nanoscale; research study; single molecule; structural biology; success; tool

DESCRIPTION (provided by applicant): We have developed a versatile technique for ultra-high-precision laser machining that uses tightly focused femtosecond laser pulses to ablate sharply defined nanometer-scale regions in materials. With this technology it is possible to selectively ablate regions even smaller than 20 nm across. This milestone achievement holds great promise for a wide range of applications, perhaps none more exciting than as a tool to highly selectively destroy intracellular structures. The ability to create structural "knockouts" in which intracellular components are selectively destroyed holds enormous promise for elucidating structure-function relationships, just as molecular-genetic knockouts have been crucial to understanding the function of genes and the proteins they encode. Toward fulfilling this potential, this proposal has dual goals of demonstrating the utility of this approach to the broader biological community, and addressing fundamental questions concerning cell division. To these ends we propose to apply structural knockout technology to study the biomechanics of the cytoskeleton and mitosis. These experiments will characterize: 1) the mechanical and force generating properties that allow chromosomes to bind and move along microtubules, 2) the antimitotic activity of the chemotherapy drug taxol, and 3) the role of centrioles in establishing mitotic architecture. The ultimate limits of structural knockout technology will also be investigated to explore the potential for intriguing future applications such as targeted disruption of single molecules or genes.

描述（由申请人提供）：我们开发了一种用于超高精度激光加工的通用技术，该技术使用紧密聚焦的飞秒激光脉冲来烧蚀材料中清晰定义的纳米级区域。利用这项技术，可以选择性地烧蚀宽度甚至小于 20 nm 的区域。这一里程碑式的成就为广泛的应用带来了巨大的希望，也许没有什么比作为高度选择性破坏细胞内结构的工具更令人兴奋的了。创建结构“敲除”（选择性破坏细胞内成分）的能力为阐明结构与功能关系带来了巨大希望，就像分子遗传敲除对于理解基因及其编码蛋白质的功能至关重要一样。为了实现这一潜力，该提案具有双重目标：向更广泛的生物界展示这种方法的实用性，并解决有关细胞分裂的基本问题。为此，我们建议应用结构敲除技术来研究细胞骨架和有丝分裂的生物力学。这些实验将表征：1）允许染色体结合并沿着微管移动的机械和力生成特性，2）化疗药物紫杉醇的抗有丝分裂活性，以及​​3）中心粒在建立有丝分裂结构中的作用。还将研究结构敲除技术的最终极限，以探索未来应用的潜力，例如单分子或基因的定向破坏。