Sub-3Å proteasome image-shift data collection¶

Instruments¶

SEMC@NYSBC Krios

1. Scope: FEI Titan-Krios, 300 kV, micro probe at low mag, nano probe at high mags
2. Camera: Gatan K2. mount below Falcon/Ceta. 1.21 Å at 29kx scope nominal mag

Grid¶

• 1.2/1.3 C-flat carbon support; 400 mesh Copper grid.

Krios1-K2-NYSBC

Application:¶

MSI-T2 Advanced (= MSI-T + "Beam Tilt Image" for coma-free alignment + "N2 Filling" for Krios auto nitrogen filler)

Presets Manager:¶

No preset cycling (Krios has its own normalization).

Queuing usage:¶

Hole Targeting: Queuing, target at the carbon area to cover four holes.
Exposure Targeting: No Queuing. automated targeting. See Exposure Targeting Set-up for MSi-T for Four Holes

Move method to reach the targets:¶

Hole: Navigator Iterative movement: Target Tolerance: 1e-7 m; Acceptable Tolerance: 3e-7 m
Focus: Presets Manager image shift
Exposure: Presets Manager image shift

Wait time before final exposure:
3 seconds
10 seconds addition for the first target¶