Nanofabricated Tools for Single Molecule Studies

用于单分子研究的纳米制造工具

• 批准号: 6643431
• 负责人: Robert C. Dunn
• 金额: $ 25.38万
• 依托单位: UNIVERSITY OF KANSAS LAWRENCE
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-09-01 至 2006-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6643431
• 关键词: atomic force microscopy; bioengineering /biomedical engineering; biomedical equipment development; calcium channel; fluorescence resonance energy transfer; image enhancement; nanotechnology; nuclear membrane; single cell analysis; technology /technique development

DESCRIPTION (provided by applicant): The technique development proposed here will result in new optical microscopy methods that have sub-20 nm spatial resolution, single molecule detection limits, Angstrom level sectioning capabilities, simultaneous fluorescence and force sensitivity, and the ability to carry out all of these measurements on living cellular systems under physiological conditions. The goal is to develop new techniques and methods that combine attributes from near-field scanning optical microscopy (NSOM), atomic force microscopy (AFM), and fluorescence resonance energy transfer (FRET) to enhance imaging capabilities for single molecule studies viable biological tissue. The NSOM/AFM and NSOM/FRET probes to be developed in specific aims 1 and 2 will be compatible with many commercial AFM instruments, adding important new contrast mechanisms for those currently conducting AFM measurements. Once tested and validated on control samples, the nanotechnology developed under specific aims 1 and 2 will be applied to studies on the nuclear membrane in specific aim 3. The high resolution and sensitivity of these techniques will be utilized to unlock key outstanding issues concerning the role and identity of the "plug" observed in nuclear pore complexes, the plug response to lumenal calcium depletion, the distribution and correlation of nuclear pores with calcium channels in the membrane, and the dynamics of plug motion in the nuclear pore complex. These experiments will be done on viable nuclear membranes under physiological conditions, something heretofore not possible with the near-field technique

描述（由申请人提供）：这里提出的技术发展将导致新的光学显微镜方法，具有低于20纳米的空间分辨率，单分子检测限，埃级切片能力，同时荧光和力灵敏度，以及在生理条件下对活细胞系统进行所有这些测量的能力。目标是开发结合近场扫描光学显微镜（NSOM），原子力显微镜（AFM）和荧光共振能量转移（FRET）属性的新技术和方法，以增强单分子研究可行生物组织的成像能力。在具体目标1和2中开发的NSOM/AFM和NSOM/FRET探针将与许多商用AFM仪器兼容，为目前进行AFM测量的人增加重要的新对比机制。一旦在对照样品上进行测试和验证，在特定目标1和2下开发的纳米技术将应用于特定目标3的核膜研究。这些技术的高分辨率和高灵敏度将用于解决核孔复合物中观察到的“堵塞物”的作用和身份，管腔钙耗尽的堵塞物响应，膜中核孔与钙通道的分布和相关性，以及核孔复合物中堵塞物运动的动力学等关键问题。这些实验将在生理条件下的活核膜上进行，这是近场技术迄今无法做到的