HeLa cells FIB milled

Description

Technical info

Tilt Series Date: 2017-03-27

Data Taken By: Shrawan kumar Mageswaran

Species / Specimen: Unspecified

Strain: None

Tilt Series Settings: Single Axis, tilt range: (-60.0°, 60.0°), step: 2°, constant angular increment, dosage: 100.0 eV/Ų, defocus: -10.0 μm, magnification: 22500x.

Microscope: Caltech Polara

Acquisition Software: SerialEM

Upload Method: reupload_downloaded

Processing Software Used: IMOD

Purification / Growth Conditions / Treatment: HeLa cells were grown on Quantifoil Au R2/2 grids in DMEM + 10% FBS + Pen/Strep from ~1-2 cells per grid square. Cells were synchronized using double thymidine block and stained with Tubulin Tracker before plunge freezing.

Sample Preparation: Cells were manually blotted from behind for ~3 sec. Cells were sputter coated using ~2 nm Pt and lamellae were milled to ~200nm thickness from different parts of the cell (not just midbodies, which was the intended experimental target).

Attached Files

TS6.mrc, Tilt Series (Pixel Size 0.48 nm), 1.4 GB

TS6_4bin_DB.rec, Reconstruction (Pixel Size 0.0 nm), 208.0 MB