MRI: Development of Programmable Substrates for Quantitative Investigation of Mechanotransduction using Holographic Optical Tweezer (HOT) Arrays

该奖项旨在将全息光镊（HOT）开发成定量研究细胞中机械传导的仪器。该仪器将是唯一能够量化的结构和动态的细胞骨架与荧光散斑显微镜（FSM），而生化信号和机械力同时控制与HOT。旋转圆盘共聚焦显微镜将为FSM提供细胞骨架的图像，而可编程闭环HOT系统将同时在4-D中操作多达约100个胶体颗粒。这些颗粒将被许多不同的生化分子功能化，并将以任何期望的模式附着到细胞膜上。同时捕获多个珠子将使纳米级力应用于单个细胞-比现有的光学方法增加多达两个数量级。闭环控制系统将能够产生巨大的力，并且在新的低级特征识别和全息计算代码的帮助下，它还将能够在真实的时间内控制和测量多个位置的力-分辨率在30 Hz时可达到10 fN。研究人员将与学术和工业合作伙伴密切合作，以确保所提出的技术被广泛采用。这个跨学科项目将汇集来自工程和生物学的学生和博士后。通过拟议的研究过程中，他们将开发胶体科学，光学，分子生物学和细胞生物学的广泛技能。这种跨学科的培训将为他们在物理和生物科学的交叉点纳米科学和纳米技术的创新职业做好准备。此外，拟议的仪器将用于物理和生物科学研究生的光学显微镜高级课程。研究人员将继续鼓励代表性不足的群体的成员为他们的研究做出贡献。

This is an award to develop Holographic Optical Tweezers (HOT) into a instrument for quantitative investigation of mechanotransduction in cells. This instrument will be uniquely capable of quantifying the structure and dynamics of the cytoskeleton with fluorescent speckle microscopy (FSM) while biochemical signals and mechanical forces are simultaneously controlled with HOT. A spinning-disc confocal microscope will provide images of the cytoskeleton for FSM, while a programmable closed-loop HOT system will simultaneously manipulate up to about one hundred colloidal particles in 4-D. These particles will be functionalized with a number of different biochemical molecules and will be attached to the cell membrane in any desired pattern. Simultaneous trapping of multiple beads will enable the application of nanonewton level forces to single cells - an increase of up to two orders of magnitude over existing optical methods. The closed-loop control system will be capable of generating large forces and, with the help of novel low-level feature recognition and hologram calculation codes, it will also be capable of controlling and measuring forces at multiple locations in real time - with resolutions as fine as 10 fN at 30Hz.The investigators will work closely with academic and industrial partners to ensure that the proposed technology is widely adopted. This interdisciplinary project will bring together students and post-docs from engineering and biology. Through the course of the proposed research, they will develop a broad set of skills in colloid science, optics, molecular biology and cell biology. This interdisciplinary training will prepare them for innovative careers in nanoscience and nanotechnology at the intersection of the physical and biological sciences. Furthermore, the proposed instrument will be employed in an advanced course in optical microscopy for graduate students in the physical and biological sciences. The investigators will continue to encourage members of underrepresented groups to contribute to their research.