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

这一奖项是为了将全息光学镊子（HOT）发展成为一种定量研究细胞机械转导的仪器。该仪器将具有独特的能力，可以用荧光散斑显微镜（FSM）定量细胞骨架的结构和动力学，同时用HOT控制生化信号和机械力。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100个胶体颗粒。这些颗粒将与许多不同的生化分子功能化，并以任何所需的模式附着在细胞膜上。同时捕获多个珠子将使纳米级的力应用于单个细胞——比现有的光学方法增加了两个数量级。闭环控制系统将能够产生巨大的力，并且在新颖的低水平特征识别和全息图计算代码的帮助下，它还将能够实时控制和测量多个位置的力，分辨率可达30Hz的10 fN。研究人员将与学术界和工业界合作伙伴密切合作，以确保拟议的技术得到广泛采用。这个跨学科的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，他们将在胶体科学，光学，分子生物学和细胞生物学方面发展广泛的技能。这种跨学科的培训将为他们在纳米科学和纳米技术领域的创新职业生涯做好准备，在物理科学和生物科学的交叉领域。此外，该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.