In this paper, robust tools for montage parallel array cryo-tomography (MPACT) tailored for vitrified specimens are presented, which can image large fields of view while preserving high-resolution structural information. The combination of multiple techniques in MPACT supports studies that contextually define the three-dimensional architecture of cells, and it provides flexibility and efficient data processing and analysis.
Imaging large fields of view while preserving high-resolution structural information remains a challenge in low-dose cryo-electron tomography. Here we present robust tools for montage parallel array cryo-tomography (MPACT) tailored for vitrified specimens. The combination of correlative cryo-fluorescence microscopy, focused-ion-beam milling, substrate micropatterning, and MPACT supports studies that contextually define the three-dimensional architecture of cells. To further extend the flexibility of MPACT, tilt series may be processed in their entirety or as individual tiles suitable for sub-tomogram averaging, enabling efficient data processing and analysis.
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