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