Rapid routine structure determination of macromolecular assemblies using electron microscopy: current progress and further challenges.
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.
Carragher
Bridget
B
Department of Cell Biology, The Scripps Research Institute, La Jolla, CA 92037, USA.
Fellmann
Denis
D
Guerra
Francisco
F
Milligan
Ronald A
RA
Mouche
Fabrice
F
Pulokas
James
J
Sheehan
Brian
B
Quispe
Joel
J
Suloway
Christian
C
Zhu
Yuanxin
Y
Potter
Clinton S
CS
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
D001704:Biopolymers; D046911:Macromolecular Substances; D006433:Hemocyanins