The cryo-electron microscopy reconstruction problem is to find the three-dimensional (3D) structure of a macromolecule given noisy samples of its two-dimensional projection images at unknown random directions. Present algorithms for finding an initial 3D structure model are based on the “angular reconstitution” method in which a coordinate system is established from three projections, and the orientation of the particle giving rise to each image is deduced from common lines among the images. However, a reliable detection of common lines is difficult due to the low signal-to-noise ratio of the images. In this paper we describe two algorithms for finding the unknown imaging directions of all projections by minimizing global self-consistency errors. In the first algorithm, the minimizer is obtained by computing the three largest eigenvectors of a specially designed symmetric matrix derived from the common lines, while the second algorithm is based on semidefinite programming (SDP). Compared with existing algorithms, the advantages of our algorithms are five-fold: first, they accurately estimate all orientations at very low common-line detection rates; second, they are extremely fast, as they involve only the computation of a few top eigenvectors or a sparse SDP; third, they are nonsequential and use the information in all common lines at once; fourth, they are amenable to a rigorous mathematical analysis using spectral analysis and random matrix theory; and finally, the algorithms are optimal in the sense that they reach the information theoretic Shannon bound up to a constant for an idealized probabilistic model.

Recently popularized randomized methods for principal component analysis (PCA) efficiently and reliably produce nearly optimal accuracy -even on parallel processors -unlike the classical (deterministic) alternatives. We adapt one of these randomized methods for use with data sets that are too large to be stored in random-access memory (RAM). (The traditional terminology is that our procedure works efficiently out-of-core.) We illustrate the performance of the algorithm via several numerical examples. For example, we report on the PCA of a data set stored on disk that is so large that less than a hundredth of it can fit in our computer's RAM.

We introduce a graph Laplacian-based algorithm for the tomographic reconstruction of a planar object from its projections taken at random unknown directions. A Laplace-type operator is constructed on the data set of projections, and the eigenvectors of this operator reveal the projection orientations. The algorithm is shown to successfully reconstruct the Shepp-Logan phantom from its noisy projections. Such a reconstruction algorithm is desirable for the structuring of certain biological proteins using cryo-electron microscopy.

Two-dimensional analysis of projections of single particles acquired by an electron microscope is a useful tool to help identifying the different kinds of projections present in a dataset and their different projection directions. Such analysis is also useful to distinguish between different kinds of particles or different particle conformations. In this paper we introduce a new algorithm for performing twodimensional multireference alignment and classification that is based on a Hierarchical clustering approach using correntropy (instead of the more traditional correlation) and a modified criterion for the definition of the clusters specially suited for cases in which the Signal-to-Noise Ratio of the differences between classes is low. We show that our algorithm offers an improved sensitivity over current methods in use for distinguishing between different projection orientations and different particle conformations. This algorithm is publicly available through the software package Xmipp.