Guidelines for using Bsoft for high resolution reconstruction and validation of biomolecular structures from electron micrographs

摘要: Cryo-electron microscopy (cryoEM) is becoming popular as a tool to solve biomolecular structures with the recent availability of direct electron detectors allowing automated acquisition high resolution data. The Bsoft software package, developed over 20 years for analyzing micrographs, offers full workflow validated single particle analysis extensive functionality, enabling customization specific cases. With increasing use cryoEM and its automation, proper validation results bigger concern. three major approaches, independent data sets, resolution-limited processing, coherence testing, can be incorporated into any workflow. Here, main divided four phases: (i) micrograph preprocessing, (ii) picking, (iii) alignment reconstruction, (iv) interpretation. Each these phases represents conceptual unit that automated, followed by check point assess results. aim in first reconstruct one or more maps at best possible. Map interpretation then involves identification components, segmentation, quantification, modeling. algorithms are well established, future plans focused on ease use, automation institutionalizing validation.