Automated image acquisition for single-particle reconstruction using p97 as the biological sample

We have used Leginon, a fully automatic system capable of acquiring cryo-el ectron micrographs, to collect data of single particles, specifically of th e AAA ATPase p97. The images were acquired under low-dose conditions and re quired no operator intervention other than the initial setup and periodic r efilling of the cold-stage dewar. Each image was acquired at two different defocus values. Two-dimensional projection maps of p97 were calculated from these data and compared to results previously obtained using the conventio nal manual data collection methods to film. The results demonstrate that Le ginon performs as well as an experienced microscopist for the acquisition o f single-particle data. The general advantages of automation are discussed. (C) 2001 Academic Press.