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

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


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.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Image Processing, Computer-Assisted*
  • Microscopy, Electron*
  • Models, Molecular
  • Molecular Structure
  • Software*
  • User-Computer Interface