Nanofabricated Tools for Single Molecule Studies

用于单分子研究的纳米制造工具

• 批准号: 6800947
• 负责人: Robert C. Dunn
• 金额: $ 5.4万
• 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/6800947
• 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）：此处提出的技术开发将导致新的光学显微镜方法具有低于20 nm的空间分辨率，单分子检测极限，Angstrom级分段能力，同时荧光和力敏感性以及在生理系统下对生物细胞系统进行所有这些测量的能力。目的是开发新技术和方法，这些技术和方法结合了近场扫描光学显微镜（NSOM），原子力显微镜（AFM）和荧光共振能传递（FRET），以增强单分子研究可爱生物学组织的成像能力。在特定目标1和2中开发的NSOM/AFM和NSOM/FRET探针将与许多商业AFM仪器兼容，为当前进行AFM测量的人添加了重要的新对比度机制。 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在膜中，以及核孔复合物中插头运动的动力学。这些实验将在生理条件下在可行的核膜上进行，这是近场技术不可能的