Agro_AD1484

Description

Technical info

Tilt Series Date: 2015-10-26

Data Taken By: Debnath Ghosal

Species / Specimen: Agrobacterium tumefaciens

Strain:

Tilt Series Settings: Single Axis, tilt range: (-60.0°, 60.0°), step: 1.5°, constant angular increment, dosage: 160.0 eV/Ų, defocus: -8.0 μm, magnification: 26000x.

Microscope: Janelia Titan

Acquisition Software: UCSFTomo

Upload Method: pipeline

Processing Software Used: Raptor

Collaborators and Roles: Strain constructed by Anath Das. Data collected and processed by Debnath Ghosal using Caltech tomography pipeline. This data was collected at the Janelia Krios.

Purification / Growth Conditions / Treatment: Agro_AD1484 (AD1484:A348dB/pAD2079,virB on a multicopy plasmid). Cells were scraped off the induction plate (particularly from the immediate surface of the plate: this is to ensure that if something is happening on the real surface of the plate, we do not miss it), dissolved in induction media, spun for 15 min in a table top centrifuge, supernatant thrown away, cells redissolved in PBS. Add gold to this: 1ul of gold to 4 ul of cells and freeze.

Sample Preparation: Vitrobot (MK iv) settings: 100% humidity, 4µl sample, Blot force 6, blot time 3 sec. Wait time 0 sec. Drain time 1 sec. Ethane/propane mix. Grid: Quantifoil Cu 2/2 200.

Attached Files

20151026_AD1484_30017_full.rec, Reconstruction (Pixel Size 1.68 nm), 430.6 MB

20151026_AD1484_30017.mrc, Tilt Series (Pixel Size 0.42 nm), 2.2 GB

dga2015-10-26-86_slicer19257.png, small irregular shaped REZ . This cell seems to be in an intermediate state. (perhaps transitioning between a state with and without a condensed nucleoid) . flow lines are visible . -andrew, 448.9 kB