wt_perfusion_fixed_immunofluoro

Tilt Series Date: 2015-10-08 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: 175.0 eV/Ų, defocus: -6.0 μm, magnification: 34000x. Microscope: Caltech Polara Acquisition Software: UCSFTomo Upload Method: pipeline Processing Software Used: Raptor Purification / Growth Conditions / Treatment: This sample is wt cells grown in perfusion set-up. It was imaged on LM for wires, fixed with 4 % formaldehyde for 1 hour, immunofluorescence was done for cytochromes, reimaged, fixed with 2.5% glutaraldehyde for 15 minutes, triple washed with distilled water and saved in distilled water until freezing. The grid was prepared by Sahand on 10/06/15. The grid was frozen on 10/06/15 at USC using manual plunger. Sample Preparation: This is gold grid with honey C. The grid was pulled out quickly from media. 1.5ul 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.