Robotic Dispensing of Cubic Phase for Crystallizing Membrane Proteins

机器人和人类增强计划建议#：308078标题：用于结晶膜蛋白的立方相机器人分配PI：郑，元俄亥俄州立大学Fdn这项研究介绍了能够自动处理纳升体积的高粘性生物材料的新的机器人技术和系统。在2001年宣布对人类基因组进行测序之后，蛋白质组学领域迫切需要机器人系统。这一里程碑式的事件导致了结构生物学领域的激烈活动，目的是确定由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，而X射线结晶学是确定结构的唯一可靠方法。最近介绍了一种使用脂质立方中间相(在介观中)来制备几种重要的膜蛋白和蛋白质复合体的衍射级晶体的方法。In MESO方法要求以高通量的方式控制和可重复地处理特定形状的珍贵、高粘度蛋白质/脂肪立方相混合物的精确、纳升体积。将使用以下三种方法来促进这种系统的开发：a)有效地规划分配工具以有效地输送粘性材料，b)利用立方相进行坐标测量，以及c)计算机视觉验证成功分配。对于高效的运动规划，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。为了定位容器的底面以进行准确的分配，PI将使用立方相本身进行坐标测量，以消除对昂贵和复杂的坐标测量机的需要。计算机视觉验证为监控输送提供了一种经济有效的方法，因为当粘性物质拒绝离开分配工具时，偶尔会发生故障。这项研究的智力价值在于创造了新的机器人技术和系统，用于自动处理以前从未研究过的纳升体积的高粘性生物材料。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发展将是这次活动的主要智力优势。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，该系统将能够每天对数千种结晶条件进行筛选，以更好地了解膜蛋白的结构；b)其结果将为自动处理通用粘性材料创建一个知识库；以及c)该教育计划寻求在包括工程学和生物化学的跨学科领域培训学生。

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.