Bdellovibrio/E. coli minicell

Description

Technical info

Tilt Series Date: 2013-12-06

Data Taken By: Yi-Wei Chang

Species / Specimen: Bdellovibrio bacteriovorus

Strain: HD100

Tilt Series Settings: Single Axis, tilt range: (-65.0°, 65.0°), step: 1.0°, constant angular increment, dosage: 120.0 eV/Ų, defocus: -8.0 μm, magnification: 26000x.

Microscope: Janelia Titan

Acquisition Software: UCSFTomo

Upload Method: pipeline

Processing Software Used: Raptor

Collaborators and Roles: Bdellovibrio Strain from Liz Sockett, WM3433 E. coli minicell from Jun Liu & William Margolin, Sample prepared by Lee Rettberg

Purification / Growth Conditions / Treatment: Bdellovibrio cells grown in S17-1 prey solution at 30 degrees, then filtered by 0.45 um filter, concentrated by centrifugation. WM3433 E. coli minicells were enriched by 2 step centrifugation (3,000 rcf, 3 min --> 9,000 rcf, 5 min). Invasion complex were made by mixing them for 36 min. Pellet of 10 nm gold beads were first resuspent thoroughly and 20 ul cells were then gently mixed in by tapping the tube

Sample Preparation: Vitrobot settings: 100% humidity, 3µl sample, 10 nm BSA treated gold. -3 offset, 4 sec blot time, no drain. Ethane/propane mix. Grid: QF Cu R2/2 200

ycw2013-12-06-32_slicer7119.jpg: nucleoid ycw2013-12-06-32_slicer7121.jpg: filament

Attached Files

montage_25x250024.mrc, Tilt Series (Pixel Size 0.42 nm), 3.6 GB

montage_25x250024_full.rec, Reconstruction (Pixel Size 1.68 nm), 688.9 MB