S. pombe (Cdc25-22)

Description

Technical info

Tilt Series Date: 2013-09-23

Data Taken By: Matt Swulius

Species / Specimen: Schizosaccharomyces pombe

Strain: cdc25-22 rlc1-3GFP

Tilt Series Settings: Single Axis, tilt range: (-55.0°, 55.0°), step: 1°, constant angular increment, dosage: 180.0 eV/Ų, defocus: -10.0 μm, magnification: 18000x.

Microscope: Caltech Polara

Acquisition Software: UCSFTomo

Upload Method: pipeline

Processing Software Used: IMOD

Collaborators and Roles: Mohan Balasubramanian and Mithilesh Mishra

Purification / Growth Conditions / Treatment: Cells were grown to OD = 0.2 and them temperature shifted to the nonpermissive temp of 36C for 4 hours. Then cells were shifted to the permissive temp of 25C and monitored by fLM for ring formation (between 30-60 min). ~90 are synchronized.

Sample Preparation: After synchronization cells were pelleted, mixed to include ~ 20% dextran and high pressure frozen. They were then cryosectioned at 150-200 nm.

Attached Files

092313_pombe0004.mrc, Tilt Series (Pixel Size 0.616 nm), 3.2 GB

092313_pombe0004.rec, Reconstruction (Pixel Size 2.464 nm), 226.7 MB