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Initial evaluation of a direct detection device detector for single particle cryo-electron microscopy |
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| Authors: | Milazzo Anna-Clare Cheng Anchi Moeller Arne Lyumkis Dmitry Jacovetty Erica Polukas James Ellisman Mark H Xuong Nguyen-Huu Carragher Bridget Potter Clinton S |
| Institution: | aDepartment of Chemistry and Biochemistry, University of California, San Diego, La Jolla, CA 92093, USA;bThe National Resource for Automated Molecular Microscopy, Department of Cell Biology, The Scripps Research Institute, La Jolla, CA 92037, USA;cThe National Center for Microscopy and Imaging Research, Center for Research on Biological Systems, University of California, San Diego, La Jolla, CA 92093, USA;dDepartment of Neurosciences, University of California, San Diego, La Jolla, CA 92093, USA |
| 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 200 keV and a nominal magnification of 29,000×. This camera has a significantly improved signal to noise ratio and modulation transfer function (MTF) at 200 keV 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 ∼30 K 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 ∼70 K GroEL particles acquired using the DDD; the quality of the density map demonstrates the potential of this new recording device for cryoEM data acquisition. |
| Keywords: | TEM Cryo-electron microscopy Direct detection device Active pixel sensor CMOS devices |
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