Author: Boyington, Jeffrey C.; Joyce, M. Gordon; Sastry, Mallika; Stewart-Jones, Guillaume B. E.; Chen, Man; Kong, Wing-Pui; Ngwuta, Joan O.; Thomas, Paul V.; Tsybovsky, Yaroslav; Yang, Yongping; Zhang, Baoshan; Chen, Lei; Druz, Aliaksandr; Georgiev, Ivelin S.; Ko, Kiyoon; Zhou, Tongqing; Mascola, John R.; Graham, Barney S.; Kwong, Peter D.
Title: Structure-Based Design of Head-Only Fusion Glycoprotein Immunogens for Respiratory Syncytial Virus Document date: 2016_7_27
ID: 1nbocmux_18
Snippet: Samples were diluted with a buffer containing 20 mM HEPES, pH 7.0, and 150 mM NaCl, adsorbed to a freshly glow-discharged carbon-film grid for 15 s, washed with the above buffer, and stained with 0.7% uranyl formate. Images were collected at a magnification of 100,000 semi-automatically using SerialEM [28] on a FEI Tecnai T20 microscope operating at 200 kV and equipped with a 2k x 2k Eagle CCD camera. The pixel size was 0.22 nm/px. Particles were.....
Document: Samples were diluted with a buffer containing 20 mM HEPES, pH 7.0, and 150 mM NaCl, adsorbed to a freshly glow-discharged carbon-film grid for 15 s, washed with the above buffer, and stained with 0.7% uranyl formate. Images were collected at a magnification of 100,000 semi-automatically using SerialEM [28] on a FEI Tecnai T20 microscope operating at 200 kV and equipped with a 2k x 2k Eagle CCD camera. The pixel size was 0.22 nm/px. Particles were picked manually or using the swarm mode in e2boxer from the EMAN2 software package [29] . The datasets for i-693, i-693 + D25 complex, i-693 + motavizumab complex, i-693 + D25 + motavizumab complex, i-447, i-447 + D25 complex, i-447 + motavizumab complex, and i-447 + D25 + motavizumab complex contained 4872, 8915, 6354, 5847, 8806, 2242, 3031, and 6542 particles, respectively. The datasets for i-273, i-273 + D25 complex, i-273 + motavizumab complex, i-273 + D25 + motavizumab complex, i-210, i-210 + D25 complex, i-210 + motavizumab complex, and i-210 + D25 + motavizumab complex contained 3652, 2840, 4017, 4169, 2584, 2123, 1341, and 1939 particles, respectively. Initial reference-free 2D classification was performed using EMAN2. In cases where the quality of 2D classes was insufficient, classification was repeated using reference-free rotational alignment and classification ("AP C") or referencefree alignment and correspondence analysis in SPIDER [30] .
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