Bdellovibrio/E. coli minicell

Description

Technical info

Tilt Series Date: 2013-09-28

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: 100.0 eV/Ų, defocus: -8.0 μm, magnification: 27500x.

Microscope: Caltech Polara

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 15 min.

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 finder

ycw2013-09-28-10_slicer10289.jpg: Bdellovibrio bacteriovorus . nucleoid . OMV . flagellum ycw2013-09-28-10_slicer14214.jpg: flagellum sheath peeling off of flagellum ycw2013-09-28-10_slicer14215.jpg: pink: end of a flagellum . green: blebbing outer membrabe? . blue: flagellar sheath peeling off of flagellum

Attached Files

20130928_YWC034_10010.mrc, Tilt Series (Pixel Size 0.39 nm), 3.7 GB

20130928_YWC034_10010_full.rec, Reconstruction (Pixel Size 1.56 nm), 712.7 MB