title: Initial evaluation of a direct detection device detector for single particle cryo-electron microscopy.
abstract: We report on initial results of using a new direct detection device (DDD) for single particle reconstruction of vitreous ice embedded specimens. Images were acquired on a Tecnai F20 at 200keV and a nominal magnification of 29,000×. This camera has a significantly improved signal to noise ratio and modulation transfer function (MTF) at 200keV compared to a standard CCD camera installed on the same microscope. Control of the DDD has been integrated into Leginon, an automated data collection system. Using GroEL as a test specimen, we obtained images of ∼30K particles with the CCD and the DDD from the same specimen sample using essentially identical imaging conditions. Comparison of the maps reconstructed from the CCD images and the DDD images demonstrates the improved performance of the DDD. We also obtained a 3D reconstruction from ∼70K GroEL particles acquired using the DDD; the quality of the density map demonstrates the potential of this new recording device for cryoEM data acquisition.
authors.affiliation: Department of Chemistry and Biochemistry, University of California, San Diego, La Jolla, CA 92093, USA.
authors.forename: Anna-Clare
authors.initials: AC
authors.lastname: Milazzo
authors.forename: Anchi
authors.initials: A
authors.lastname: Cheng
authors.forename: Arne
authors.initials: A
authors.lastname: Moeller
authors.forename: Dmitry
authors.initials: D
authors.lastname: Lyumkis
authors.forename: Erica
authors.initials: E
authors.lastname: Jacovetty
authors.forename: James
authors.initials: J
authors.lastname: Polukas
authors.forename: Mark H
authors.initials: MH
authors.lastname: Ellisman
authors.forename: Nguyen-Huu
authors.initials: NH
authors.lastname: Xuong
authors.forename: Bridget
authors.initials: B
authors.lastname: Carragher
authors.forename: Clinton S
authors.initials: CS
authors.lastname: Potter
mesh_terms: D002244:Carbon / Q000737:chemistry; D018834:Chaperonin 60 / Q000737:chemistry; D020285:Cryoelectron Microscopy / Q000295:instrumentation* / Q000379:methods*; D005583:Fourier Analysis; D007091:Image Processing, Computer-Assisted / Q000295:instrumentation*; D059629:Signal-To-Noise Ratio
chemical_list: D018834:Chaperonin 60; D002244:Carbon