Laser Tweezer Based Instrument for Cell Biology Research

描述（由申请人提供）： 拟议的项目是开发一种细胞生物学研究仪器，其基础是一种以可持续的亚微米分辨率扰动关键信号转导介质的局部细胞溶质浓度的技术。在这种技术中，微球用选定的酶修饰，在颗粒表面产生高局部浓度的效应物。激光镊子被用来操纵细胞中的这些粒子。第一阶段项目将证明基于波形调制实现光学捕获系统的可行性，并使用相对低成本的二极管激光器代替前期工作中使用的钛蓝宝石激光器来证明该技术的有效性。这项技术的成功应用将产生广泛的影响。最重要的是，它将提供一种在更大的时间尺度上影响局部胞质性质的方法（例如，小时），这比细胞生物学家目前使用诸如局灶性撑开和电生理学技术等技术所能实现的要多，由于扩散，这两种技术通常在毫秒至秒内导致效应物的广泛细胞分布。所提出的仪器将提供直接将生物化学、药效团或所选蛋白质递送到细胞内任何位点的能力，从而构成生物医学研究领域的巨大技术进步。

DESCRIPTION (provided by applicant): The proposed project is to develop an instrument for cell biology research based on a technique for perturbing local cytosolic concentrations of key signal transduction mediators with sustainable submicrometer resolution. In this technique, microspheres are decorated with selected enzymes, creating high local concentrations of an effector at the particle surface. Laser tweezers are used to manipulate these particles in a cell. The Phase I project will prove the feasibility of implementing the optical trapping system based on waveform modulation and demonstrate the effectiveness of the technique using a relatively low-cost diode laser in place of the titanium sapphire laser used in preliminary work. Successful application of the technology will have broad implications. Most importantly, it would provide a means for effecting local cytosolic properties over timescales much greater (e.g., hours) than cell biologists currently are able to achieve using techniques such as focal uncaging and electrophysiological techniques, both of which typically result in broad cellular distribution of effectors within milliseconds to seconds due to diffusion. The proposed instrument will provide the capability to directly deliver a biochemical, pharmacophore or protein of choice to any site within a cell and thereby constitutes a tremendous technological advance for the biomedical research field.