wt_perfusion_fixed_immunofluoro

Description

Technical info

Tilt Series Date: 2015-10-26

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: 160.0 eV/Ų, defocus: -5.0 μm, magnification: 34000x.

Microscope: Caltech Polara

Acquisition Software: UCSFTomo

Upload Method: pipeline

Processing Software Used: Raptor

Collaborators and Roles: Moh and Sahand USC

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 10/22/15. The grid was frozen on 10/23/15 at Caltech using vitrobot MK4.

Sample Preparation: The sample was frozen using vitrobot. C side left. 1.8 ul gold 10nm size was added on opposite side while on vitrobot. conditions: blot force=6, blot time=2.5s, drain time =1s, wait time=0. humidity set 80%. temp 22C. Use 2.5ul gold next time

Attached Files

wt_immunofluorocyto_sq60007_full.rec, Reconstruction (Pixel Size 1.28 nm), 534.5 MB

wt_immunofluorocyto_sq60007.mrc, Tilt Series (Pixel Size 0.32 nm), 3.4 GB