- EMDB-5336: Unprocessed map of GroEL obtained with a CCD camera to compare re... -
+
データを開く
IDまたはキーワード:
読み込み中...
-
基本情報
登録情報
データベース: EMDB / ID: EMD-5336
タイトル
Unprocessed map of GroEL obtained with a CCD camera to compare results from cryo SPA with a DDD camera using the same grid. Initial evaluation of a Direct Detection Device detector for single particle cryo-electron microscopy.
マップデータ
This is a map from approx. 30K particles of GroEL, unfiltered and not amplitude corrected using a CCD camera.
ジャーナル: J Struct Biol / 年: 2011 タイトル: Initial evaluation of a direct detection device detector for single particle cryo-electron microscopy. 著者: Anna-Clare Milazzo / Anchi Cheng / Arne Moeller / Dmitry Lyumkis / Erica Jacovetty / James Polukas / Mark H Ellisman / Nguyen-Huu Xuong / Bridget Carragher / Clinton S Potter / 要旨: 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 200keV and a ...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 200keV and a nominal magnification of 29,000×. This camera has a significantly improved signal to noise ratio and modulation transfer function (MTF) at 200keV 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 ∼30K 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 ∼70K GroEL particles acquired using the DDD; the quality of the density map demonstrates the potential of this new recording device for cryoEM data acquisition.
詳細: Protochips C-flat grid, holey carbon with 2um holes and 2um spacing 400 mesh copper grid
凍結
凍結剤: ETHANE / チャンバー内湿度: 100 % / チャンバー内温度: 93 K / 装置: OTHER 詳細: Vitrification instrument: Vitrobot. Grid plasma cleaned for 20 seconds with Fischione 1020 plasma cleaner using 75 percent argon, 25 percent oxygen mix. 手法: Temperature of chamber was 4 degrees C. 0 seconds drain time. Single blot. 0 mm offset. 4 uL sample applied to grid. Blot for 3.5 seconds before plunging.
-
電子顕微鏡法
顕微鏡
FEI TECNAI F20
温度
最低: 102 K / 最高: 102 K / 平均: 102 K
アライメント法
Legacy - 非点収差: The objective lens astigmatism was corrected automatically using Leginon Legacy - Electron beam tilt params: beam tilt was less than 0.1 mrad