Robotic Dispensing of Cubic Phase for Crystallizing Membrane Proteins

机器人和人类扩增程序摘要提案号：308078标题：机器人分配立方相结晶膜蛋白PI：郑，袁俄亥俄州立大学研究部本研究地址新的机器人技术和系统，使纳升体积的高粘度生物材料的自动化处理。 在2001年宣布人类基因组测序完成后，蛋白质组学界迫切需要机器人系统。 这一具有里程碑意义的事件导致了结构生物学的激烈活动，旨在确定基因组编码的蛋白质的三维结构。 这些蛋白质中约有三分之一是细胞膜的组成部分，X射线晶体学是确定结构的唯一可靠方法。 最近已经引入了一种使用脂质立方中间相（在meso中）的方法，用于生产几种重要的膜蛋白和蛋白质复合物的衍射质量晶体。 内消旋方法需要以高通量的方式对精确的、纳升体积的具有限定形状的珍贵的、高粘性的蛋白质/脂质立方相混合物进行受控的和可再现的处理。 三种方法将被用来促进这样一个系统的发展，如下：a）有效的运动规划的分配工具，有效地提供粘性材料，B）利用立方相的坐标测量，和c）计算机视觉验证成功的分配。 为了有效的运动规划，PI将研究分配工具相对于接收容器的运动轨迹，以确保在预定形状的纳升范围内输送准确的立方相体积。 为了定位容器的底部表面以进行精确分配，PI将使用立方相本身进行坐标测量，以避免对昂贵且复杂的坐标测量机的需求。 计算机视觉验证提供了一种具有成本效益的方式来监控交付，因为故障可能会偶尔发生时，粘性材料拒绝离开点胶tool.The智力的优点，这项研究是在创造新的机器人技术和系统的自动处理纳升体积的高粘性生物材料，从来没有被研究过。与运动规划、坐标测量和计算机视觉检测相关的三项新技术的开发将是这项活动的主要智力价值。 这项研究的广泛影响包括：a）该项目将产生一个功能齐全的机器人系统，该系统将能够每天筛选数千种结晶条件，以加快对膜蛋白结构的更好理解，B）结果将为通用粘性材料的自动处理创建知识库，以及c）教育计划旨在培养学生在工程和生物化学等跨学科领域的能力。

Robotics and Human Augmentation ProgramABSTRACTProposal #: 308078Title: Robotic Dispensing of Cubic Phase for Crystallizing Membrane ProteinsPI: Zheng, YuanOhio State Univ Res FdnThis research addresses new robotic technologies and systems which enable the automated handling of nanoliter volumes of highly viscous bio-materials. The robotic system is urgently needed by the proteomics community following on the announcement in 2001 that the human genome had been sequenced. This landmark event has led to intense activity in structural biology aimed at determining the 3-dimensional structure of the proteins coded for by the genome. About a third of these proteins are integral to the cell's membranes and x-ray crystallography is the only reliable method for structure determination. A method that uses the lipid cubic mesophase (in meso) has recently been introduced for producing diffraction quality crystals of several important membrane proteins and a protein complex. The in meso method requires the controlled and reproducible handling in high-throughput fashion of accurate, nanoliter volumes of precious, highly viscous protein/lipid cubic phase mixtures of defined shape. Three approaches will be used to facilitate the development of such a system as follows: a) efficient-motion planning of the dispensing tool for effective delivery of viscous materials, b) utilization of the cubic phase for coordinate measuring, and c) computer vision verification of successful dispensing. For efficient-motion planning the PIs will study the motion trajectory of the dispensing tool with respect to the receiving container to assure delivery of accurate cubic phase volumes in the nanoliter range of predetermined shape. To locate the bottom surface of the container for accurate dispensing, the PIs will use the cubic phase itself for coordinate measurement to obviate the need for expensive and complex coordinate measuring machines. Computer vision verification provides a cost effective way to monitor delivery since failure may occur occasionally when the viscous material refuses to leave the dispensing tool.The intellectual merits of this research are in the creation of new robotic technologies and systems for the automatic handling of nanoliter volumes of highly viscous bio-materials which have never been studied before. The development of the three new technologies related to motion planning, coordinates measuring, and computer-vision inspection will be the primary intellectual merits of this activity. The broader impacts of this research include the following: a) the project will produce a fully functional robotic system that will enable the screening of thousands of crystallization conditions daily to expedite the better understanding of the structure of membrane proteins, b) the results will create a knowledge base for the automatic handling of generic viscous materials, and c) the educational plan seeks to train students in an interdisciplinary area encompassing engineering and biochemistry.