|Title||SPIDER image processing for single-particle reconstruction of biological macromolecules from electron micrographs.|
|Publication Type||Journal Article|
|Year of Publication||2008|
|Authors||Shaikh, TR, Gao, H, Baxter, B, Asturias, FJ, Boisset, N, Leith, AD, Frank, J|
|Keywords||Image Processing, Computer-Assisted, Microscopy, Electron, Models, Molecular, Molecular Structure, Software, User-Computer Interface|
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.
|Alternate Journal||Nat Protoc|
|PubMed Central ID||PMC2737740|