Operator-assisted harvesting of protein crystals using a universal micromanipulation robot.
Operator-assisted harvesting of protein crystals using a universal micromanipulation robot.
复制标题
使用通用微型手持机器人对蛋白质晶体的辅助收集。
DOI：
10.1107/s0021889807012149
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发表时间：
2007-06-01
影响因子：
6.1
通讯作者：
Rupp, Bernhard
中科院分区：
文献类型：
--
作者：
Viola, Robert;Carman, Peter;Walsh, Jace;Miller, Echo;Benning, Matthew;Frankel, Daniel;McPherson, Alexander;Cudney, Bob;Rupp, Bernhard
介绍了一种用于晶体收获的通用微操作机器人的原型，机器人收获的胰酶晶体产生了高分辨率的结构，证明了机器人收获蛋白质晶体的可行性。高通量结晶学已经达到自动化水平，完整的计算机辅助机器人结晶管道能够进行鸡尾酒准备、结晶板设置以及结果检查和解释。虽然晶体引脚的安装、数据收集和结构解决方案是高度自动化的，但晶体采集和冷冻冷却仍然是实现全自动化的艰巨挑战。为了解决实现全自动化高通量结晶学的最终前沿，设计并测试了一台拟人六轴通用微操作机器人的原型，该机器人能够由操作员辅助从各种96孔板上收获和冷冻淬灭小到10 微米的蛋白质晶体。UMR配备了多功能工具交换器，提供了完全的操作灵活性。用超磁共振仪采集和冷冻冷却的胰酶晶体产生了1.5 ä的结构，证明了机器人采集蛋白质晶体的可行性。
The prototype of a universal micromanipulation robot for crystal harvesting is presented, and a robotically harvested trypsin crystal yields a high-resolution structure demonstrating the feasibility of robotic protein crystal harvesting. High-throughput crystallography has reached a level of automation where complete computer-assisted robotic crystallization pipelines are capable of cocktail preparation, crystallization plate setup, and inspection and interpretation of results. While mounting of crystal pins, data collection and structure solution are highly automated, crystal harvesting and cryocooling remain formidable challenges towards full automation. To address the final frontier in achieving fully automated high-throughput crystallography, the prototype of an anthropomorphic six-axis universal micromanipulation robot (UMR) has been designed and tested; this UMR is capable of operator-assisted harvesting and cryoquenching of protein crystals as small as 10 µm from a variety of 96-well plates. The UMR is equipped with a versatile tool exchanger providing full operational flexibility. Trypsin crystals harvested and cryoquenched using the UMR have yielded a 1.5 Å structure demonstrating the feasibility of robotic protein crystal harvesting.