HeLa (Myo1a KD) - PM damaged

Description

Technical info

Tilt Series Date: 2018-06-16

Data Taken By: Shrawan kumar Mageswaran

Species / Specimen: HeLa cells

Strain: ATCC

Tilt Series Settings: Single Axis, tilt range: (-60.0°, 60.0°), step: 2°, constant angular increment, dosage: 100.0 eV/Ų, defocus: -8.0 μm, magnification: 26000x.

Microscope: Caltech Titan Krios

Acquisition Software: SerialEM

Upload Method: pipeline

Processing Software Used: IMOD and tomo3D

Collaborators and Roles: HeLa cell line courtesy of Max Planck Institute (MPI). Wei Yang grew cells on grids, transfected with siRNA and caused plasma membrane damage using Laser before freezing. Shrawan performed CLEM and ECT.

Purification / Growth Conditions / Treatment: Cells grown on R2/2 quantifoil gold grids treated with 0.1 mg/mL fibronectin.

Sample Preparation: Cells were knocked down for Myo1a with SMART pool siRNA for 48 hr (2 x 24 hr). Laser - 640nm, 100%, 35 cycles, speed 7, Damaged area - 1.5umx1.5um. Cells were treated with 1.3uM AlPcS3a ~10 min before damage.

Attached Files

TS3.mrc, Tilt Series (Pixel Size 0.541 nm), 1.6 GB