γ\gamma-SUP: A clustering algorithm for cryo-electron microscopy images of asymmetric particles

Cryo-electron microscopy (cryo-EM) has recently emerged as a powerful tool for obtaining three-dimensional (3D) structures of biological macromolecules in native states. A minimum cryo-EM image data set for deriving a meaningful reconstruction is comprised of thousands of randomly orientated projections of identical particles photographed with a small number of electrons. The computation of 3D structure from 2D projections requires clustering, which aims to enhance the signal to noise ratio in each view by grouping similarly oriented images. Nevertheless, the prevailing clustering techniques are often compromised by three characteristics of cryo-EM data: high noise content, high dimensionality and large number of clusters. Moreover, since clustering requires registering images of similar orientation into the same pixel coordinates by 2D alignment, it is desired that the clustering algorithm can label misaligned images as outliers. Herein, we introduce a clustering algorithm $\gamma$-SUP to model the data with a $q$-Gaussian mixture and adopt the minimum $\gamma$-divergence for estimation, and then use a self-updating procedure to obtain the numerical solution. We apply $\gamma$-SUP to the cryo-EM images of two benchmark macromolecules, RNA polymerase II and ribosome. In the former case, simulated images were chosen to decouple clustering from alignment to demonstrate $\gamma$-SUP is more robust to misalignment outliers than the existing clustering methods used in the cryo-EM community. In the latter case, the clustering of real cryo-EM data by our $\gamma$-SUP method eliminates noise in many views to reveal true structure features of ribosome at the projection level.Comment: Published in at http://dx.doi.org/10.1214/13-AOAS680 the Annals of Applied Statistics (http://www.imstat.org/aoas/) by the Institute of Mathematical Statistics (http://www.imstat.org

An Imprint of Molecular Cloud Magnetization in the Morphology of the Dust Polarized Emission

We describe a morphological imprint of magnetization found when considering the relative orientation of the magnetic field direction with respect to the density structures in simulated turbulent molecular clouds. This imprint was found using the Histogram of Relative Orientations (HRO): a new technique that utilizes the gradient to characterize the directionality of density and column density structures on multiple scales. We present results of the HRO analysis in three models of molecular clouds in which the initial magnetic field strength is varied, but an identical initial turbulent velocity field is introduced, which subsequently decays. The HRO analysis was applied to the simulated data cubes and mock-observations of the simulations produced by integrating the data cube along particular lines of sight. In the 3D analysis we describe the relative orientation of the magnetic field $\mathbf{B}$ with respect to the density structures, showing that: 1.The magnetic field shows a preferential orientation parallel to most of the density structures in the three simulated cubes. 2.The relative orientation changes from parallel to perpendicular in regions with density over a critical density $n_{T}$ in the highest magnetization case. 3.The change of relative orientation is largest for the highest magnetization and decreases in lower magnetization cases. This change in the relative orientation is also present in the projected maps. In conjunction with simulations HROs can be used to establish a link between the observed morphology in polarization maps and the physics included in simulations of molecular clouds.Comment: (16 pages, 11 figures, submitted to ApJ 05MAR2013, accepted 07JUL2013

Unsupervised cryo-EM data clustering through adaptively constrained K-means algorithm

In single-particle cryo-electron microscopy (cryo-EM), K-means clustering algorithm is widely used in unsupervised 2D classification of projection images of biological macromolecules. 3D ab initio reconstruction requires accurate unsupervised classification in order to separate molecular projections of distinct orientations. Due to background noise in single-particle images and uncertainty of molecular orientations, traditional K-means clustering algorithm may classify images into wrong classes and produce classes with a large variation in membership. Overcoming these limitations requires further development on clustering algorithms for cryo-EM data analysis. We propose a novel unsupervised data clustering method building upon the traditional K-means algorithm. By introducing an adaptive constraint term in the objective function, our algorithm not only avoids a large variation in class sizes but also produces more accurate data clustering. Applications of this approach to both simulated and experimental cryo-EM data demonstrate that our algorithm is a significantly improved alterative to the traditional K-means algorithm in single-particle cryo-EM analysis.Comment: 35 pages, 14 figure

GENFIRE: A generalized Fourier iterative reconstruction algorithm for high-resolution 3D imaging

Tomography has made a radical impact on diverse fields ranging from the study of 3D atomic arrangements in matter to the study of human health in medicine. Despite its very diverse applications, the core of tomography remains the same, that is, a mathematical method must be implemented to reconstruct the 3D structure of an object from a number of 2D projections. In many scientific applications, however, the number of projections that can be measured is limited due to geometric constraints, tolerable radiation dose and/or acquisition speed. Thus it becomes an important problem to obtain the best-possible reconstruction from a limited number of projections. Here, we present the mathematical implementation of a tomographic algorithm, termed GENeralized Fourier Iterative REconstruction (GENFIRE). By iterating between real and reciprocal space, GENFIRE searches for a global solution that is concurrently consistent with the measured data and general physical constraints. The algorithm requires minimal human intervention and also incorporates angular refinement to reduce the tilt angle error. We demonstrate that GENFIRE can produce superior results relative to several other popular tomographic reconstruction techniques by numerical simulations, and by experimentally by reconstructing the 3D structure of a porous material and a frozen-hydrated marine cyanobacterium. Equipped with a graphical user interface, GENFIRE is freely available from our website and is expected to find broad applications across different disciplines.Comment: 18 pages, 6 figure

Calipso: Physics-based Image and Video Editing through CAD Model Proxies

We present Calipso, an interactive method for editing images and videos in a physically-coherent manner. Our main idea is to realize physics-based manipulations by running a full physics simulation on proxy geometries given by non-rigidly aligned CAD models. Running these simulations allows us to apply new, unseen forces to move or deform selected objects, change physical parameters such as mass or elasticity, or even add entire new objects that interact with the rest of the underlying scene. In Calipso, the user makes edits directly in 3D; these edits are processed by the simulation and then transfered to the target 2D content using shape-to-image correspondences in a photo-realistic rendering process. To align the CAD models, we introduce an efficient CAD-to-image alignment procedure that jointly minimizes for rigid and non-rigid alignment while preserving the high-level structure of the input shape. Moreover, the user can choose to exploit image flow to estimate scene motion, producing coherent physical behavior with ambient dynamics. We demonstrate Calipso's physics-based editing on a wide range of examples producing myriad physical behavior while preserving geometric and visual consistency.Comment: 11 page

Direct 3D Tomographic Reconstruction and Phase-Retrieval of Far-Field Coherent Diffraction Patterns

We present an alternative numerical reconstruction algorithm for direct tomographic reconstruction of a sample refractive indices from the measured intensities of its far-field coherent diffraction patterns. We formulate the well-known phase-retrieval problem in ptychography in a tomographic framework which allows for simultaneous reconstruction of the illumination function and the sample refractive indices in three dimensions. Our iterative reconstruction algorithm is based on the Levenberg-Marquardt algorithm. We demonstrate the performance of our proposed method with simulation studies