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TitleElectron-event representation data enable efficient cryoEM file storage with full preservation of spatial and temporal resolution.
Journal, issue, pagesIUCrJ, Vol. 7, Issue Pt 5, Page 860-869, Year 2020
Publish dateSep 1, 2020
AuthorsHui Guo / Erik Franken / Yuchen Deng / Samir Benlekbir / Garbi Singla Lezcano / Bart Janssen / Lingbo Yu / Zev A Ripstein / Yong Zi Tan / John L Rubinstein /
PubMed AbstractDirect detector device (DDD) cameras have revolutionized electron cryomicroscopy (cryoEM) with their high detective quantum efficiency (DQE) and output of movie data. A high ratio of camera frame ...Direct detector device (DDD) cameras have revolutionized electron cryomicroscopy (cryoEM) with their high detective quantum efficiency (DQE) and output of movie data. A high ratio of camera frame rate (frames per second) to camera exposure rate (electrons per pixel per second) allows electron counting, which further improves the DQE and enables the recording of super-resolution information. Movie output also allows the correction of specimen movement and compensation for radiation damage. However, these movies come at the cost of producing large volumes of data. It is common practice to sum groups of successive camera frames to reduce the final frame rate, and therefore the file size, to one suitable for storage and image processing. This reduction in the temporal resolution of the camera requires decisions to be made during data acquisition that may result in the loss of information that could have been advantageous during image analysis. Here, experimental analysis of a new electron-event representation (EER) data format for electron-counting DDD movies is presented, which is enabled by new hardware developed by Thermo Fisher Scientific for their Falcon DDD cameras. This format enables the recording of DDD movies at the raw camera frame rate without sacrificing either spatial or temporal resolution. Experimental data demonstrate that the method retains super-resolution information and allows the correction of specimen movement at the physical frame rate of the camera while maintaining manageable file sizes. The EER format will enable the development of new methods that can utilize the full spatial and temporal resolution of DDD cameras.
External linksIUCrJ / PubMed:32939278 / PubMed Central
MethodsEM (single particle)
Resolution2.07 - 3.3 Å
Structure data

EMDB-22346:
CryoEM structure of human light chain apoferritin calculated from EER movies (super-resolution experiment, no supersampling)
Method: EM (single particle) / Resolution: 3.3 Å

EMDB-22347:
CryoEM structure of human light chain apoferritin calculated from EER movies (super-resolution experiment, 2x supersampling with random subpixel information)
Method: EM (single particle) / Resolution: 2.8 Å

EMDB-22348:
CryoEM structure of human light chain apoferritin calculated from EER movies (super-resolution experiment, 2x supersampling)
Method: EM (single particle) / Resolution: 2.4 Å

EMDB-22349:
CryoEM structure of human light chain apoferritin calculated from EER movies (intra-fraction motion correction experiment, no supersampling)
Method: EM (single particle) / Resolution: 2.14 Å

EMDB-22350:
CryoEM structure of human light chain apoferritin calculated from EER movies (intra-fraction motion correction experiment, 1.5x supersampling, particle motion corrected without B-spline interpolation
Method: EM (single particle) / Resolution: 2.1 Å

EMDB-22351:
CryoEM structure of human light chain apoferritin calculated from EER movies (intra-fraction motion correction experiment, 1.5x supersampling, particle motion corrected with B-spline interpolation)
Method: EM (single particle) / Resolution: 2.07 Å

Source
  • Homo sapiens (human)

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