wt_perfusion_fixed

Tilt Series Date: 2015-09-19 Data Taken By: Poorna Subramanian Species / Specimen: Shewanella oneidensis Strain: MR-1 Tilt Series Settings: Single Axis, tilt range: (-60.0°, 60.0°), step: 1.0°, constant angular increment, dosage: 135.0 eV/Ų, defocus: -6.0 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.倾斜序列日期：2015-09-19 数据采集者：Poorna Subramanian 物种/标本：奥奈达希瓦氏菌（Shewanella oneidensis） 菌株：MR-1 倾斜序列设置：单轴倾斜模式，倾斜范围为(-60.0°, 60.0°)，步长1.0°，采用恒定角度增量，电子剂量135.0 eV/Ų，欠焦值-6.0 μm，放大倍数41000× 显微镜：加州理工学院Polara电镜（Caltech Polara） 采集软件：UCSFTomo 上传方式：流水线（pipeline） 所用处理软件：镜检下细胞状态良好。下次实验建议使用1.5 μL金颗粒 合作者及分工：Moh与Sahand，南加州大学（USC） 纯化/培养条件与处理方式：本样品为野生型（wild type，wt）细胞，采用灌流装置培养，经2.5%戊二醛固定15分钟。载网由Sahand于2015年9月10日制备：先后用蒸馏水三次漂洗，随后将载网保存于蒸馏水中，直至2015年9月15日在南加州大学使用手动 plunger 进行 plunge 冷冻 样品制备：本样品采用带有honey C膜的金载网。从培养基中快速取出载网后，立即向载网的C面（即细胞附着的一侧）加入1.2 μL 10 nm金颗粒。将载网置于手动 plunger 上，从C面的对侧（背面）进行滤纸吸附，随后进行 plunge 冷冻