title: Rapid routine structure determination of macromolecular assemblies using electron microscopy: current progress and further challenges.
abstract: Although the methodology of molecular microscopy has enormous potential, it is time consuming and labor intensive. The techniques required to produce a three-dimensional (3D) electron density map of a macromolecular structure normally require manual operation of an electron microscope by a skilled operator and manual supervision of the sometimes complex software needed for analysis and calculation of 3D maps. Systems to automate the process of data acquisition from an electron microscope are being developing and these systems are being integrated with specimen handling operations and post acquisition data processing. Here, the current performance of our existing systems and the future challenges involved in substantially improving both the sustained throughput and the yield of automated data collection and analysis are reported.
authors.affiliation: Department of Cell Biology, The Scripps Research Institute, La Jolla, CA 92037, USA.
authors.forename: Bridget
authors.initials: B
authors.lastname: Carragher
authors.forename: Denis
authors.initials: D
authors.lastname: Fellmann
authors.forename: Francisco
authors.initials: F
authors.lastname: Guerra
authors.forename: Ronald A
authors.initials: RA
authors.lastname: Milligan
authors.forename: Fabrice
authors.initials: F
authors.lastname: Mouche
authors.forename: James
authors.initials: J
authors.lastname: Pulokas
authors.forename: Brian
authors.initials: B
authors.lastname: Sheehan
authors.forename: Joel
authors.initials: J
authors.lastname: Quispe
authors.forename: Christian
authors.initials: C
authors.lastname: Suloway
authors.forename: Yuanxin
authors.initials: Y
authors.lastname: Zhu
authors.forename: Clinton S
authors.initials: CS
authors.lastname: Potter
mesh_terms: D000465:Algorithms; D001704:Biopolymers / Q000737:chemistry*; D017798:Comovirus / Q000648:ultrastructure; D019544:Equipment Failure Analysis; D005240:Feasibility Studies; D006433:Hemocyanins / Q000648:ultrastructure; D007089:Image Enhancement / Q000295:instrumentation / Q000379:methods*; D007090:Image Interpretation, Computer-Assisted / Q000295:instrumentation / Q000379:methods*; D021621:Imaging, Three-Dimensional / Q000295:instrumentation / Q000379:methods*; D046911:Macromolecular Substances; D008854:Microscopy, Electron / Q000295:instrumentation* / Q000379:methods*; D008958:Models, Molecular; D008968:Molecular Conformation; D012371:Robotics / Q000295:instrumentation* / Q000379:methods*; D013048:Specimen Handling / Q000295:instrumentation / Q000379:methods; D018511:Systems Integration; D014027:Tobacco Mosaic Virus / Q000648:ultrastructure
chemical_list: D001704:Biopolymers; D046911:Macromolecular Substances; D006433:Hemocyanins