title: The effect of overabundant projection directions on 3D reconstruction algorithms.
abstract: The experimental process of collecting images from macromolecules in an electron microscope is such that it does not allow for prior specification of the angular distribution of the projection images. As a consequence, an uneven distribution of projection directions may occur. Concerns have been raised recently about the behavior of 3D reconstruction algorithms for the case of unevenly distributed projections. It has been illustrated on experimental data that in the case of a heavily uneven distribution of projection directions some algorithms tend to elongate the reconstructed volumes along the overloaded direction so much as to make a quantitative biological analysis impossible. In answer to these concerns we have developed a strategy for quantitative comparison and optimization of 3D reconstruction algorithms. We apply this strategy to quantitatively analyze algebraic reconstruction techniques (ART) with blobs, simultaneous iterative reconstruction techniques (SIRT) with voxels, and weighted backprojection (WBP). We show that the elongation artifacts that had been previously reported can be strongly reduced. With our specific choices for the free parameters of the three algorithms, WBP reconstructions tend to be inferior to those obtained with either SIRT or ART and the results obtained with ART are comparable to those with SIRT, but at a very small fraction of the computational cost of SIRT.
authors.affiliation: Centro Nacional de Biotecnología (CSIC), Universidad Autónoma Madrid, Madrid, 28049, Spain.
authors.forename: C O
authors.initials: CO
authors.lastname: Sorzano
authors.forename: R
authors.initials: R
authors.lastname: Marabini
authors.forename: N
authors.initials: N
authors.lastname: Boisset
authors.forename: E
authors.initials: E
authors.lastname: Rietzel
authors.forename: R
authors.initials: R
authors.lastname: Schröder
authors.forename: G T
authors.initials: GT
authors.lastname: Herman
authors.forename: J M
authors.initials: JM
authors.lastname: Carazo
mesh_terms: D000465:Algorithms; D000818:Animals; D020285:Cryoelectron Microscopy / Q000379:methods; D006454:Hemoglobins / Q000737:chemistry; D021621:Imaging, Three-Dimensional / Q000295:instrumentation / Q000379:methods* / Q000592:standards; D008432:Mathematical Computing; D009835:Oligochaeta / Q000737:chemistry; D011487:Protein Conformation
chemical_list: D006454:Hemoglobins