Cryo-EM reconstruction of the chlororibosome to 3.2 Å resolution within 24 h

Forsberg BO, Aibara S, Kimanius D, Paul B, Lindahl E, Amunts A

Int Union Crystallogr J IUCr J Int Union Cryst J IUCrJ 4 (6) 723-727 [2017-11-01; online 2017-09-22]

The introduction of direct detectors and the automation of data collection in cryo-EM have led to a surge in data, creating new opportunities for advancing computational processing. In particular, on-the-fly workflows that connect data collection with three-dimensional reconstruction would be valuable for more efficient use of cryo-EM and its application as a sample-screening tool. Here, accelerated on-the-fly analysis is reported with optimized organization of the data-processing tools, image acquisition and particle alignment that make it possible to reconstruct the three-dimensional density of the 70S chlororibosome to 3.2 Å resolution within 24 h of tissue harvesting. It is also shown that it is possible to achieve even faster processing at comparable quality by imposing some limits to data use, as illustrated by a 3.7 Å resolution map that was obtained in only 80 min on a desktop computer. These on-the-fly methods can be employed as an assessment of data quality from small samples and extended to high-throughput approaches.

Cryo-EM [Service]

PubMed 29123673

DOI 10.1107/s205225251701226x

Crossref 10.1107/s205225251701226x

pmc: PMC5668856
pii: kf5004

Publications 9.5.0