Evaluation of super-resolution performance of the K2 electron-counting camera using 2D crystals of aquaporin-0

Po-Lin Chiu; Xueming Li; Zongli Li; Brian Beckett; Axel F Brilot; Nikolaus Grigorieff; David A Agard; Yifan Cheng; Thomas Walz

doi:10.1016/j.jsb.2015.08.015

Evaluation of super-resolution performance of the K2 electron-counting camera using 2D crystals of aquaporin-0

J Struct Biol. 2015 Nov;192(2):163-73. doi: 10.1016/j.jsb.2015.08.015. Epub 2015 Aug 28.

Authors

Po-Lin Chiu¹, Xueming Li², Zongli Li³, Brian Beckett⁴, Axel F Brilot⁴, Nikolaus Grigorieff⁵, David A Agard⁶, Yifan Cheng⁷, Thomas Walz⁸

Affiliations

¹ Department of Cell Biology, Harvard Medical School, 240 Longwood Avenue, Boston, MA 02115, USA.
² The Keck Advanced Microscopy Laboratory, Department of Biochemistry and Biophysics, University of California San Francisco, 600 16th Street, San Francisco, CA 94158, USA.
³ Department of Cell Biology, Harvard Medical School, 240 Longwood Avenue, Boston, MA 02115, USA; The Howard Hughes Medical Institute, Harvard Medical School, 240 Longwood Avenue, Boston, MA 02115, USA.
⁴ Department of Biochemistry, Rosenstiel Basic Medical Sciences Research Center, Brandeis University, 415 South Street, Waltham, MA 02454, USA.
⁵ Janelia Research Campus, 19700 Helix Drive, Ashburn, VA 20147, USA.
⁶ The Keck Advanced Microscopy Laboratory, Department of Biochemistry and Biophysics, University of California San Francisco, 600 16th Street, San Francisco, CA 94158, USA; The Howard Hughes Medical Institute, University of California San Francisco, 600 16th Street, San Francisco, CA 94158, USA.
⁷ The Keck Advanced Microscopy Laboratory, Department of Biochemistry and Biophysics, University of California San Francisco, 600 16th Street, San Francisco, CA 94158, USA. Electronic address: ycheng@ucsf.edu.
⁸ Department of Cell Biology, Harvard Medical School, 240 Longwood Avenue, Boston, MA 02115, USA; The Howard Hughes Medical Institute, Harvard Medical School, 240 Longwood Avenue, Boston, MA 02115, USA. Electronic address: twalz@hms.harvard.edu.

Abstract

The K2 Summit camera was initially the only commercially available direct electron detection camera that was optimized for high-speed counting of primary electrons and was also the only one that implemented centroiding so that the resolution of the camera can be extended beyond the Nyquist limit set by the physical pixel size. In this study, we used well-characterized two-dimensional crystals of the membrane protein aquaporin-0 to characterize the performance of the camera below and beyond the physical Nyquist limit and to measure the influence of electron dose rate on image amplitudes and phases.

Keywords: Direct electron detection device camera; Electron counting; Super-resolution; Two-dimensional crystal.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

Animals
Aquaporins / chemistry*
Cryoelectron Microscopy / instrumentation
Cryoelectron Microscopy / methods*
Crystallization
Crystallography, X-Ray
Electrons
Eye Proteins / chemistry*
Image Processing, Computer-Assisted / methods*
Lens Capsule, Crystalline*
Limit of Detection
Membrane Proteins / analysis*
Sheep

Substances

Aquaporins
Eye Proteins
Membrane Proteins
aquaporin 0

Abstract

Publication types

MeSH terms

Substances

Grants and funding