Compares a set of particles with the corresponding projections of a reference volume. The set of particles must have a 3D angular assignment. This protocol refines the CTF, computing local defocus change. The maximun allowed defocus is a parameter introduced by the user (advanced). The protocol gives back the input set of particles with the refine local defocus and the defocus change with relation to the global defocus.
This algorithm addresses the initial volume problem in SPA by setting it in a Weighted Least Squares framework and calculating the weights through a statistical approach based on the cumulative density function of different image similarity measures.
Computes an initial 3d model from a set of projections/classes using RANSAC algorithm. This method is based on an initial non-lineal dimensionality reduction approach which allows to select representative small sets of class average images capturing the most of the structural information of the particle under study. These reduced sets are then used to generate volumes from random orientation assignments. The best volume is determined from these guesses using a random sample consensus (RANSAC) approach.
This protocol wraps *e2initialmodel.py* EMAN2 program. It will take a set of class-averages/projections and build a set of 3-D models suitable for use as initial models in single particle reconstruction. The output set is theoretically sorted in order of quality (best one is numbered 1), though it's best to look at the other answers as well. See more details in: http://blake.bcm.edu/emanwiki/EMAN2/Programs/e2initialmodel
ETHAN is a program for automatic detection of spherical particles from electron micrographs. The ETHAN software was written at the Department of Computer Science of University of Helsinki, Finland by Teemu Kivioja.
This protocol runs Relion autopicking (version > 3.0). This Relion protocol uses the 'relion_autopick' program to pick particles from micrographs, either using references (2D averages or 3D volumes) The picking with this protocol is divided in three steps: 1) Run with 'Optimize' option for several (less than 30) micrographs. 2) Execute the wizard to refine the picking parameters. 3) Run with 'Pick all' option to pick particles from all micrographs. The first steps will use internally the option '--write-fom-maps' to write to disk the FOM maps. The expensive part of this calculation is to calculate a probability-based figure-of-merit (related to the cross-correlation coefficient between each rotated reference and all positions in the micrographs. That's why it is only done in an small subset of the micrographs, where one should use representative micrographs for the entire data set, e.g. a high and a low-defocus one, and/or with thin or thick ice. Step 2 uses a much cheaper peak-detection algorithm that uses the threshold and minimum distance parameters.
This protocol wraps SPIDER CL HC command. Finds clusters of images/elements in factor space (or a selected subspace) by using Diday's method of moving centers, and applies hierarchical ascendant classification (HAC) (using Ward's method) to the resulting cluster centers.
Wrapper to CryoSparc 2D clustering program. Classify particles into multiple 2D classes to facilitate stack cleaning and removal of junk particles. Also useful as a sanity check to investigate particle quality.
This class contains a re-implementation to a method for the localized three-dimensional reconstruction of such subunits. After determining the particle orientations, local areas corresponding to the subunits can be extracted and treated as single particles.
This protocol wraps *e2refine_easy.py* EMAN2 program.This is the primary single particle refinement program in EMAN2.1+.It replaces earlier programs such as e2refine.py and e2refine_evenodd.py.Major features of this program: * While a range of command-line options still exist. You should not normally specify more than a few basic requirements. The rest will be auto-selected for you. * This program will split your data in half and automatically refine the halves independently to produce a gold standard resolution curve for every step in the refinement. * An HTML report file will be generated as this program runs, telling you exactly what it decided to do and why, as well as giving information about runtime, etc while the job is still running. * The gold standard FSC also permits us to automatically filter the structure at each refinement step. The resolution you specify is a target, NOT the filter resolution.
Protocol to refine a 3D map using Relion.Relion employs an empirical Bayesian approach to refinementof (multiple) 3D reconstructionsor 2D class averages in electron cryo-microscopy (cryo-EM). Manyparameters of a statistical model are learned from the data,whichleads to objective and high-quality results.
Apply non-uniform refinement to achieve higher resolution and map quality, especially for membrane proteins. Non-uniform refinement iteratively accounts for regions of a structure that have disordered or flexible density causing local loss of resolution. Accounting for these regions and dynamically estimating their locations can significantly improve resolution in other regions as well as overall map quality by impacting the alignment of particles and reducing the tendency for refinement algorithms to over-fit disordered regions.
This is a 3D refinement protocol whose main input is a volume and a set of particles. The set of particles has to be at full size (the finer sampling rate available), but the rest of inputs (reference volume and masks) can be at any downsampling factor. The protocol scales the input images and volumes to a reasonable size depending on the resolution of the previous iteration. The protocol works with any input volume, whichever its resolution, as long as it is a reasonable initial volume for the set of particles. The protocol does not resolve the heterogeneous problem (it assumes an homogeneous population), although it is somewhat tolerant through the use of particle weights in the reconstruction process. It is recommended to perform several global alignment iterations before entering into the local iterations. The switch from global to local should be performed when a substantial percentage of the particles do not move from one iteration to the next. The algorithm reports the cross correlation (global alignment) or cost (local) function per defocus group, so that we can see which was the percentile of each particle in its defocus group. You may want to perform iterations one by one, and remove from one iteration to the next, those particles that worse fit the model.
Reference-based refinement using SPIDER AP SHC and AP REF commands. Iterative refinement improves the accuracy in the determination of orientations. This improvement is accomplished by successive use of more finely-sampled reference projections. Two different workflows are suggested: with defocus groups or without (gold-standard refinement). For more information, see: [[http://spider.wadsworth.org/spider_doc/spider/docs/techs/recon/mr.html][SPIDER documentation on projection-matching]]
This protocol does local resolution estimation using Relion. This program basically performs a series of post-processing operations with a small soft, spherical mask that is moved over the entire map, while using phase-randomisation to estimate the convolution effects of that mask.