Towards In Vivo Force Spectroscopy Using Optomagnetic Tweezers

使用光磁镊子进行体内力谱分析

• 批准号: 8536341
• 负责人: ROBERT L CLARK
• 金额: $ 11.11万
• 依托单位: UNIVERSITY OF ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-01 至 2015-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8536341
• 关键词: Accounting; Affect; Atomic Force Microscopy; Automobile Driving; Biological; Caliber; Calibration; Cell Line; Cell physiology; Cells; Chemicals; Chemistry; Communities; Custom; Development; Disease; Endocytosis; Environment; Freedom; Health; Human; Implant; In Vitro; Individual; Lasers; Life; Magnetism; Malignant Neoplasms; Measurement; Measures; Mechanics; Methods; Microfabrication; Microscopic; Microspheres; Molecular; Nanosphere; Optics; Performance; Physiologic pulse; Positioning Attribute; Property; Proteins; Research; Research Personnel; Resolution; Scheme; Spectrum Analysis; Surface; System; Techniques; Torque; cell motility; design; driving force; flexibility; granulocyte; improved; in vivo; insight; instrument; laser tweezer; magnetic field; novel; particle; response; single molecule; therapy development; uptake

DESCRIPTION (provided by applicant): Single molecule force spectroscopy (SMFS) is used to study the fundamental forces that drive the biological activity of cells and proteins. Nearly al of these biophysical measurements are performed in vitro using purified biomolecules. When studying the function and mechanics of cells, this approach yields inherently incomplete results by failing to account for the influence of the intracellular environment. Therefore, the development of a force spectroscopy system capable of in vivo sensing will advance biophysical measurement capability and help clarify the origins of the interactions driving cellular function. Broadly, a more complete understanding of cellular processes will aid researchers in the discovery and development of treatments for various disorders including cancer. This proposal outlines the development of a novel instrument that combines the manipulation capabilities of optical and magnetic tweezers to facilitate high resolution positioning as well as the application of a relatively large range of forces inside a living cell. To realize an instrumen of this type, an optical tweezers system will be retrofitted with a magnetic apparatus capable of generating controllable rotating magnetic fields. Magnetically anisotropic Janus spheres, less than 200 nm in diameter, will be fabricated to serve as probes that can be optically trapped and magnetically actuated. Their size will facilitate cellular uptake via endocytosis. Once inside the cell infrared laser beams or ultrafast laser pulses combined with external magnetic actuation will enable probe motility and force application without compromising the health of the cell. Advanced servo control schemes will allow the accurate application of forces and measurement of intracellular mechanics. Initially probes will be inserted into live granulocytes to access instrument performance. Once parameters are optimized the in vivo force spectroscopy instrument will be made available to the general research community for the characterization of a myriad of cell lines.

描述（由申请人提供）：单分子力光谱（SMF）用于研究驱动细胞和蛋白质生物学活性的基本力。这些生物物理测量几乎是使用纯化的生物分子在体外进行的。在研究细胞的功能和力学时，这种方法通过无法说明细胞内环境的影响而产生固有的不完整结果。因此，能够在体内传感的力光谱系统的发展将提高生物物理测量能力，并有助于阐明驱动细胞功能的相互作用的起源。从广义上讲，对细胞过程的更全面了解将有助于研究人员发现和开发包括癌症在内的各种疾病的治疗方法。 该建议概述了一种新型仪器的开发，该仪器结合了光学和磁镊子的操纵能力，以促进高分辨率定位以及在活细胞内的相对较大的力施加。为了实现这种类型的仪器，将使用能够生成可控旋转磁场的磁性设备进行改装。直径小于200 nm的磁各向异性janus球将被制造为可以光学捕获和磁性驱动的探针。它们的大小将通过内吞作用促进细胞摄取。一旦进入细胞红外激光束或超快激光脉冲与外部磁性致动的结合将使探测运动和强力应用，而不会损害细胞的健康状况。高级伺服控制方案将允许精确应用力和细胞内力学的测量。最初，探针将插入活粒细胞中以访问仪器性能。一旦优化了参数，体内力光谱仪器将被提供给一般研究界，以表征无数的细胞系。