3D Clustering - RSORT3D: Sort out 3D heterogeneity of 2D data whose 3D reconstruction parameters (xform.projection) have been determined already using 3D sorting.

Usage in command line

sxrsort3d.py  stack  outdir  mask  --previous_run1=sort3d_run1_directory  --previous_run2=sort3d_run2_directory  --focus=3D_focus_mask  --radius=outer_radius  --delta=angular_step  --CTF  --sym=symmetry  --number_of_images_per_group=images_per_group  --nxinit=nxinit  --smallest_group=smallest_group  --chunkdir=chunkdir  --ir=inner_radius  --maxit=max_iter  --rs=ring_step  --xr=xr  --yr=yr  --ts=ts  --an=angular_neighborhood  --center=centring_method  --nassign=nassign  --nrefine=nrefine  --stoprnct=stop_percent  --function=user_function  --independent=independent_runs  --low_pass_filter=low_pass_filter  --unaccounted  --seed=random_seed  --group_size_for_unaccounted=group_size_for_unaccounted  --sausage  --PWadjustment=PWadjustment  --protein_shape=protein_shape  --upscale=upscale  --wn=wn  --interpolation=method

sxrsort3d.py exists only in MPI version.

mpirun -np 192 sxrsort3d.py bdb:data rsort3d mask.hdf --previous_run1=sort3d1 --previous_run2=sort3d2 --radius=88 --maxit=25 --independent=3 --CTF --number_of_images_per_group=11000 --low_pass_filter=.20 --chunkdir=./ --sym=c4 --PWadjustment=pwrec.txt

stack

Input images stack: (default required string)

outdir

Output directory: There is a log.txt that describes the sequences of computations in the program. (default required string)

mask

3D mask: (default none)

--previous_run1

Directory of 1st sort3d run: (default required string)

--previous_run2

Directory of 2nd sort3d run: (default required string)

--focus

Focus 3D mask: Mask used for focused clustering (default none)

--radius

Outer radius for rotational correlation [Pixels]: Must be smaller than half the box size. Please set to the radius of the particle. (default -1)

--delta

Angular step for projections: (default '2')

--CTF

Use CTF: Do a full CTF correction during the alignment. (default False)

--sym

Point-group symmetry: (default c1)

--number_of_images_per_group

Images per group: Critical number of images per group, defined by user. (default 1000)

--nxinit

Initial image size for sorting: (default 64)

--smallest_group

Smallest group size: Minimum members for identified group. (default 500)

--chunkdir

Directory containing chunk files: Typically, specify meridien output directory. A chunk file contains a list of particle IDs belonging to associated halfset. This information is used for computing margin of error. (default none)

--ir

Inner radius for rotational correlation [Pixels]: Must be bigger than 1. (default 1)

--maxit

Maximum iterations: (default 50)

--rs

Step between rings in rotational correlation: Must be bigger than 0. (default 1)

--xr

X search range [Pixels]: The translational search range in the x direction will take place in a +/xr range. (default '1')

--yr

Y search range [Pixels]: The translational search range in the y direction. If omitted it will be set as xr. (default '-1')

--ts

Translational search step [Pixels]: The search will be performed in -xr, -xr+ts, 0, xr-ts, xr, can be fractional. (default '0.25')

--an

Local angular search width [Degrees]: This defines the neighbourhood where the local angular search will be performed. (default '-1')

--center

Centering method: 0 - if you do not want the volume to be centered, 1 - center the volume using the center of gravity. (default 0)

--nassign

Number of reassignment iterations: Performed for each angular step. (default 1)

--nrefine

Number of alignment iterations: Performed for each angular step. (default 0)

--stoprnct

Assignment convergence threshold [%]: Used to asses convergence of the run. It is the minimum percentage of assignment change required to stop the run. (default 3.0)

--function

Reference preparation function: Function used to prepare the reference volume. (default do_volume_mrk05)

--independent

Number of independent runs: Number of independent equal-Kmeans(default 3)

--low_pass_filter

Low-pass filter frequency [1/Pixel]: Low-pass filter used for the 3D sorting on the original image size. (default -1.0)

--unaccounted

Reconstruct unaccounted images: (default False)

--seed

Random seed: Seed used for the initial random assignment for EQ Kmeans. The program generates a random integer by default. (default -1)

--group_size_for_unaccounted

Unaccounted particles group size: (default 500)

--sausage

Use sausage filter: (default False)

--PWadjustment

Power spectrum reference: Text file containing a 1D reference power spectrum. (default none)

--protein_shape

Protein Shape: It defines protein preferred orientation angles. “g” is for globular proteins and “f” is for filament proteins. (default g)

--upscale

Power spectrum adjustment strength: This parameters adjusts how strongly the power spectrum of the volume should be modified to match the reference. A value of 1 brings the volume's power spectrum completely to the reference, while a value of 0 means no modification. (default 0.5)

--wn

Target image size [Pixels]: If different than 0, then the images will be rescaled to fit this size. (default 0)

--interpolation

3D interpolation method: Method interpolation in 3D. Options are tr1 or 4nn. (default 4nn)

sxrsort3d.py finds out stable members by carrying out two-way comparison of two independent sxsort3d.py runs.

For small tested datasets (real and simulated ribosome data around 10K particles), it gives 70%-90% reproducibility. However, this rate also depends on the choice of number of images per group and number of particles in the smallest group.

On lonestar cluster of TACC, using 264 cpus, it takes about 2 hours and 23 minutes to accomplish 95953 128×128 images for one sxsort3d.py independent run, 2 hours 24 minutes to accomplish one independent sxrsort3d.py run with number_of_images_per_group set as 30000.

K-means, equal K-means, reproducibility, two-way comparison.

Not published yet.

Zhong Huang

Category 1:: APPLICATIONS

sparx/bin/sxrsort3d.py

sxmeridien, sxheader, sx3dvariability, sxsort3d and sxsort3d_depth

Alpha:: Under development.

There are no known bugs so far.