S. pombe (Cdc25-22)

Description

Technical info

Tilt Series Date: 2014-04-19

Data Taken By: Matt Swulius

Species / Specimen: Schizosaccharomyces pombe

Strain: cdc25-22 rlc1-3GFP

Tilt Series Settings: Single Axis, tilt range: (-60.0°, 60.0°), step: 1°, constant angular increment, dosage: 200.0 eV/Ų, defocus: -8.0 μm, magnification: 18000x.

Microscope: Caltech Polara

Acquisition Software: UCSFTomo

Upload Method: pipeline

Processing Software Used: IMOD

Collaborators and Roles: Mithilesh Mishra, Mohan Balasubramanian

Purification / Growth Conditions / Treatment: Synchronized cytokinesis. From OD 600 = 0.25 grow for 4 hrs at 36 C. Return to 25 C for 45 min then begin monitoring ring formation by LM. Once 80-90 of rings are continuous they were mixed with Dextran (100 ul of 40% Dextran in ~ 400 ul of pelleted cells). They were then high pressure frozen.

Sample Preparation: High pressure freezing followed by cryosectioning

Attached Files

041914_pombe_0006.mrc, Tilt Series (Pixel Size 0.6 nm), 3.4 GB

041914_pombe_0006.rec, Reconstruction (Pixel Size 2.4 nm), 1.1 GB