Dataset Information

Cryo-EM reconstruction of heavy chain mouse apoferritin, 8 datasets testing dose rate and correlative double sampling mode for the Gatan K3 direct electron detector. [1143 multi-frame micrographs composed of 120 frames in LZW compressed TIFF format

ABSTRACT:

SUBMITTER: Ming Sun

PROVIDER: EMPIAR-10588 | biostudies-other |

REPOSITORIES: biostudies-other

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Publications

Practical considerations for using K3 cameras in CDS mode for high-resolution and high-throughput single particle cryo-EM.

Sun Ming M Azumaya Caleigh M CM Tse Eric E Bulkley David P DP Harrington Matthew B MB Gilbert Glenn G Frost Adam A Southworth Daniel D Verba Kliment A KA Cheng Yifan Y Agard David A DA

Journal of structural biology 20210511 3

Detector technology plays a pivotal role in high-resolution and high-throughput cryo-EM structure determination. Compared with the first-generation, single-electron counting direct detection camera (Gatan K2), the latest K3 camera is faster, larger, and now offers a correlated-double sampling mode (CDS). Importantly this results in a higher DQE and improved throughput compared to its predecessor. In this study, we focused on optimizing camera data collection parameters for daily use within a cry ...[more]

PMID: 33984504

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Project description:The capture of transient scenes at high imaging speed has been long sought by photographers, with early examples being the well known recording in 1878 of a horse in motion and the 1887 photograph of a supersonic bullet. However, not until the late twentieth century were breakthroughs achieved in demonstrating ultrahigh-speed imaging (more than 10(5) frames per second). In particular, the introduction of electronic imaging sensors based on the charge-coupled device (CCD) or complementary metal-oxide-semiconductor (CMOS) technology revolutionized high-speed photography, enabling acquisition rates of up to 10(7) frames per second. Despite these sensors' widespread impact, further increasing frame rates using CCD or CMOS technology is fundamentally limited by their on-chip storage and electronic readout speed. Here we demonstrate a two-dimensional dynamic imaging technique, compressed ultrafast photography (CUP), which can capture non-repetitive time-evolving events at up to 10(11) frames per second. Compared with existing ultrafast imaging techniques, CUP has the prominent advantage of measuring an x-y-t (x, y, spatial coordinates; t, time) scene with a single camera snapshot, thereby allowing observation of transient events with temporal resolution as tens of picoseconds. Furthermore, akin to traditional photography, CUP is receive-only, and so does not need the specialized active illumination required by other single-shot ultrafast imagers. As a result, CUP can image a variety of luminescent--such as fluorescent or bioluminescent--objects. Using CUP, we visualize four fundamental physical phenomena with single laser shots only: laser pulse reflection and refraction, photon racing in two media, and faster-than-light propagation of non-information (that is, motion that appears faster than the speed of light but cannot convey information). Given CUP's capability, we expect it to find widespread applications in both fundamental and applied sciences, including biomedical research.

Dataset Information

Cryo-EM reconstruction of heavy chain mouse apoferritin, 8 datasets testing dose rate and correlative double sampling mode for the Gatan K3 direct electron detector. [1143 multi-frame micrographs composed of 120 frames in LZW compressed TIFF format

Publications

Practical considerations for using K3 cameras in CDS mode for high-resolution and high-throughput single particle cryo-EM.

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