Bdellovibrio2492/E.coli minicell

Description

Technical info

Tilt Series Date: 2014-07-18

Data Taken By: Yi-Wei Chang

Species / Specimen: Bdellovibrio bacteriovorus

Strain: HD100

Tilt Series Settings: Single Axis, tilt range: (-60.0°, 60.0°), step: 1.0°, constant angular increment, dosage: 100.0 eV/Ų, defocus: -10.0 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 HI 2492 grown from frozen stocks in liquid PY media at 37C for 7 days, subculturing half of culture into equal volume of fresh media every two days. E. coli minicells WM3433 grown O/N to high OD and spun down 7 mins at 6krpm. Supernatant spun 10 mins at 14krpm to pellet minicells. Minicells resuspended in 20ul media and combined in 1:1 v/v ratio with Bdellovibrio.

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

ycw2014-07-18-29_slicer10477.jpg: Bdellovibrio bacteriovorus HI 2492 in contact with WM3433 E. coli minicell. Bdello, marked with arrow, is not feeling so good.

Attached Files

20140718_Janelia_YWC107_10018.mrc, Tilt Series (Pixel Size 0.33 nm), 1.7 GB

20140718_Janelia_YWC107_10018_full.rec, Reconstruction (Pixel Size 1.32 nm), 688.9 MB