wt_perfusion_fixed

Tilt Series Date: 2015-09-16 Data Taken By: Poorna Subramanian Species / Specimen: Shewanella oneidensis Strain: MR-1 Tilt Series Settings: Single Axis, tilt range: (-65.0°, 65.0°), step: 1.0°, constant angular increment, dosage: 130.0 eV/Ų, defocus: -7.0 μm, magnification: 41000x. Microscope: Caltech Polara Acquisition Software: UCSFTomo Upload Method: pipeline Processing Software Used: The cells look good on scope. Next time use 1.5ul gold. Collaborators and Roles: Moh and Sahand USC Purification / Growth Conditions / Treatment: This sample is wt cells grown in perfusion set-up. It is fixed with 2.5% glutaraldehyde for 15 minutes. The grid was prepared by Sahand on 9/10/15. The grid was triple rinsed in distilled water and saved in distilled water until freezing on 9/15/15 at USC using manual plunger. Sample Preparation: This is gold grid with honey C. The grid was pulled out quickly from media. 1.2ul of 10nm gold was added immediately to the cells on the C side -same side as cells were present. The grid was loaded on manual plunger and blotted from behind- opposite of C side. And then plunge frozen.

倾斜序列采集日期：2015-09-16
数据采集者：普尔纳·苏布拉马尼亚安（Poorna Subramanian）
物种/样品：奥奈达希瓦氏菌（Shewanella oneidensis）
菌株：MR-1
倾斜序列设置：单轴倾斜，倾斜范围为(-65.0°, 65.0°)，步长1.0°，采用恒定角度增量，电子剂量为130.0 eV/Ų，欠焦量为-7.0 μm，放大倍数为41000×
显微镜：加州理工学院Polara型透射电镜
采集软件：UCSFTomo
上传方式：流水线（pipeline）
所用处理软件：本次电镜观测下细胞状态良好，建议后续实验使用1.5 μL金颗粒
合作人员及分工：莫（Moh）、萨汉德（Sahand），美国南加州大学（USC）
纯化/培养条件及处理方式：本样品为野生型细胞，采用灌流装置培养。使用2.5%戊二醛固定15分钟。样品栅格由萨汉德于2015年9月10日制备。随后用蒸馏水进行三次漂洗，并保存于蒸馏水中，直至2015年9月15日在美国南加州大学使用手动冷冻弹射器（manual plunger）完成快速冷冻制样
样品制备：本样品采用带有蜂窝碳膜（honey C）的金栅格。将栅格从培养基中快速取出后，立即向细胞所在的碳膜侧加入1.2 μL 10 nm金颗粒。将栅格安装至手动冷冻弹射器上，从碳膜侧的背面进行吸滤，随后完成快速冷冻制样