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Nat Protoc. 2008;3(12):1941-74. doi: 10.1038/nprot.2008.156.

SPIDER image processing for single-particle reconstruction of biological macromolecules from electron micrographs.

Author information

1
Wadsworth Center, Empire State Plaza, Albany, New York 12201-0509, USA.

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.

PMID:
19180078
PMCID:
PMC2737740
DOI:
10.1038/nprot.2008.156
[Indexed for MEDLINE]
Free PMC Article

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