Abstract：
:The power of cryo-electron tomography (cryoET) lies in its capability to characterize macromolecules in their cellular context. Structure determination by cryoET, however, is time-consuming compared to single particle approaches. A recent study reported significant acceleration of data acquisition by a fast-incremental single-exposure (FISE) tilt series scheme. Here we improved the method and evaluated its efficiency and performance. We show that (1) FISE combined with the latest generation of direct electron detectors speeds up collection considerably, (2) previous generation (pre-2017) double-tilt axis Titan Krios holders are also suitable for FISE data acquisition, (3) x, y and z-specimen shifts can be compensated for, and (4) FISE tilt series data can generate averages of sub-nanometer resolution. These advances will allow for a widespread adoption of cryoET for high-throughput in situ studies and high-resolution structure determination across different biological research disciplines.
journal_name
J Struct Bioljournal_title
Journal of structural biologyauthors
Eisenstein F,Danev R,Pilhofer Mdoi
10.1016/j.jsb.2019.08.006subject
Has Abstractpub_date
2019-11-01 00:00:00pages
107-114issue
2eissn
1047-8477issn
1095-8657pii
S1047-8477(19)30176-5journal_volume
208pub_type
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