Uber D C, Jaklevic J M, Theil E H, Lishanskaya A, McNeely M R
Human Genome Center, Lawrence Berkeley Laboratory, University of California, Berkeley 94720.
Biotechniques. 1991 Nov;11(5):642-7.
We describe a system that applies image processing and robotic techniques to automatically pick individual colonies from square petri dishes and array them in 96-well microtiter plates. Digital images of the colony distribution in the dishes are acquired using a video camera and frame buffer. Commercial image processing software is used to identify individual colonies and determine their locations. A Hewlett-Packard Microassay System robot reads the resulting coordinate file for each dish, picks cells from each identified colony and transfers the cells into a microtiter plate well. A disposable pipet tip is used as the sterile implement for colony picking. Custom holders position the dishes accurately and provide common coordinate systems for imaging and picking. The system is calibrated to account for the depth of agar in the dishes. The robot can process up to 10 dishes and 20 plates (1920 colonies) in a single run. It has successfully arrayed a cosmid library of the S. pombe genome consisting of approximately 6000 colonies in 30 petri dishes in about 40 hours of robot time. Future enhancements to the system are discussed.
我们描述了一种系统，该系统应用图像处理和机器人技术从方形培养皿中自动挑选单个菌落，并将它们排列在96孔微量滴定板中。使用摄像机和帧缓冲器获取培养皿中菌落分布的数字图像。使用商业图像处理软件识别单个菌落并确定其位置。惠普微量分析系统机器人读取每个培养皿生成的坐标文件，从每个识别出的菌落中挑选细胞，并将细胞转移到微量滴定板孔中。一次性移液器吸头用作菌落挑选的无菌工具。定制的支架可精确放置培养皿，并为成像和挑选提供通用坐标系统。该系统经过校准以考虑培养皿中琼脂的深度。该机器人单次运行最多可处理10个培养皿和20个平板（1920个菌落）。它已成功在约40小时的机器人操作时间内，将由约6000个菌落组成的粟酒裂殖酵母基因组黏粒文库排列在30个培养皿中。文中还讨论了该系统未来的改进方向。
