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Protocol
Nature Protocols 3, 1941–1974 (1 December 2008) | doi:10.1038/nprot.2008.156
SPIDER image processing for single-particle reconstruction of biological macromolecules from electron micrographs
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Abstract
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.
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