Correlative montage parallel array cryo-tomography for in situ structural cell biology

Yang, Jie E.; Larson, Matthew R.; Sibert, Bryan S.; Kim, Joseph Y.; Parrell, Daniel; Sanchez, Juan C. (ORCID:000000021178183X); Pappas, Victoria (ORCID:0000000239274055); Kumar, Anil; Cai, Kai; Thompson, Keith; Wright, Elizabeth R. (ORCID:0000000307129475)

doi:10.1038/s41592-023-01999-5

DOE OSTI · 2000776

Correlative montage parallel array cryo-tomography for in situ structural cell biology

Yang, Jie E.·Larson, Matthew R.·Sibert, Bryan S.·Kim, Joseph Y.·Parrell, Daniel·Sanchez, Juan C. (ORCID:000000021178183X)·Pappas, Victoria (ORCID:0000000239274055)·Kumar, Anil·Cai, Kai·Thompson, Keith·Wright, Elizabeth R. (ORCID:0000000307129475)

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Abstract

Abstract 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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59 BASIC BIOLOGICAL SCIENCES Biochemistry & Molecular Biology

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Yang, Jie E., Larson, Matthew R., Sibert, Bryan S., Kim, Joseph Y., Parrell, Daniel, Sanchez, Juan C. (ORCID:000000021178183X), Pappas, Victoria (ORCID:0000000239274055), Kumar, Anil, Cai, Kai, Thompson, Keith, Wright, Elizabeth R. (ORCID:0000000307129475). 2023-09-18. Correlative montage parallel array cryo-tomography for in situ structural cell biology. https://doi.org/10.1038/s41592-023-01999-5

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