title: SPIDER image processing for single-particle reconstruction of biological macromolecules from electron micrographs.
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.
authors.affiliation: Wadsworth Center, Empire State Plaza, Albany, New York 12201-0509, USA.
authors.forename: Tanvir R
authors.initials: TR
authors.lastname: Shaikh
authors.forename: Haixiao
authors.initials: H
authors.lastname: Gao
authors.forename: William T
authors.initials: WT
authors.lastname: Baxter
authors.forename: Francisco J
authors.initials: FJ
authors.lastname: Asturias
authors.forename: Nicolas
authors.initials: N
authors.lastname: Boisset
authors.forename: Ardean
authors.initials: A
authors.lastname: Leith
authors.forename: Joachim
authors.initials: J
authors.lastname: Frank
mesh_terms: D007091:Image Processing, Computer-Assisted; D008854:Microscopy, Electron; D008958:Models, Molecular; D015394:Molecular Structure; D012984:Software; D014584:User-Computer Interface