%0 Journal Article %J Nat Protoc %D 2008 %T SPIDER image processing for single-particle reconstruction of biological macromolecules from electron micrographs. %A Shaikh, Tanvir R %A Gao, Haixiao %A Baxter, Bill %A Asturias, Francisco J %A Boisset, Nicolas %A Leith, ArDean %A Frank, Joachim %K Image Processing, Computer-Assisted %K Microscopy, Electron %K Models, Molecular %K Molecular Structure %K Software %K User-Computer Interface %X

This protocol describes the reconstruction of biological molecules from the electron micrographs of single particles. Computation here is performed using the image-processing software SPIDER and can be managed using a graphical user interface, termed the SPIDER Reconstruction Engine. Two approaches are described to obtain an initial reconstruction: random-conical tilt and common lines. Once an existing model is available, reference-based alignment can be used, a procedure that can be iterated. Also described is supervised classification, a method to look for homogeneous subsets when multiple known conformations of the molecule may coexist.

%B Nat Protoc %V 3 %P 1941-74 %8 10/2008 %G eng %U http://www.ncbi.nlm.nih.gov/pubmed/19180078 %N 12 %R 10.1038/nprot.2008.156