reducefiltvol(1) General Commands Manual reducefiltvol(1) NAME reducefiltvol - Reduce a volume and/or filter it in Fourier space SYNOPSIS reducefiltvol [options] input_file output_file DESCRIPTION Reducefiltvol provides a wrapper script for two operations that produce tomograms with higher signal-to-noise ratio suitable for visualization of features, segmentation, and detection of particle positions: size reduction and filtering. Either one or both of these operations can be done. The reduction is done with antialias filtering by Binvol, so large reduction factors can be used without introducing noise from aliasing. The filtering is done on a 3-D Fourier transform with Mtf- filter(1). The options here provide either a simple low-pass filter to remove high frequencies, or a Wiener-like deconvolution filter that matches what is implemented in Warp (Tegunov and Cramer, 2019, Nat. Methods 16, 1146-1152) and IsoNet (Y. T. Yiu, et al., 2022, Nat. Comm. 13, 6482). The filter compensates for attenuations by the microscope contrast transfer function (CTF), using an assumed signal-to-noise ratio that starts high and falls off at a specified rate. Pixel size and origin values in the header of the output file are set so that the coordinate system matches that of the input file. Thus, models drawn on the input file in 3dmod should load correctly onto the output file, and vice versa. This coordinate system can be attached to IMOD models generated on the reduced volume by external software using the -i option to Imodtrans. OPTIONS Reducefiltvol uses the PIP package for input (see the manual page for pip). Options can be specified either as command line arguments (with the -) or one per line in a command file (without the -). Options can be abbreviated to unique letters; the currently valid abbreviations for short names are shown in parentheses. -input (-i) OR -InputFile File name Input file with volume to be reduced and/or filtered -output (-o) OR -OutputFile File name Output file for reduced and/or filtered volume -reduce (-r) OR -ReductionFactor Floating point Overall reduction factor to apply, which will be used for X and Y even if a different factor is entered for Z. -zfactor (-z) OR -ZReductionFactor Floating point Factor to reduce by in Z; the default is to reduce the same in all dimensions. Can be non-integer. -lowpass (-l) OR -LowPassRadiusSigma Two floats Cutoff radius and sigma for a low pass filter that imposes a high-frequency Gaussian roll-off to 0.0. This option cannot be entered with -deconv. -deconv (-dec) OR -DeconvolutionStrength Floating point The option activates a deconvolution filter by specifying the overall strength of the deconvolution, which controls the degree to which low frequencies are boosted. Values around 0.25 to 1 may be useful. The bigger the value, the more low frequencies are accentuated. This value is treated as it is in Warp; values used in IsoNet need to be divided by 3 to have the same effect here. This option cannot be entered with -lowpass. The following 7 entries are used for the deconvolution filter. -snr (-sn) OR -SNRFalloff Floating point Falloff rate with frequency of the signal-to-noise ratio assumed for computing the deconvolution filter; higher values attenuate high frequencies more. Some users suggest keeping this value comparable to the -deconv entry. The default is 0.7, the value in IsoNet. -dchigh (-dch) OR -HighPassNyquist Floating point Cutoff frequency of a high-pass filter to prevent excessive boosting of the lowest frequencies, where the CTF is low. For consistency with Warp and IsoNet, the value is expressed as the fraction of Nyquist, and is thus twice the usual frequency in reciprocal pixels. The default is 0.02 as in IsoNet. -pixel (-pi) OR -PixelSize Floating point Pixel size in nanometers. This entry is needed only if the pixel size in the image file header is incorrect, but the pro- gram will try to determine is value by getting the raw stack pixel size in the Ctfplotter command file and comparing pixel sizes in the raw stack and input volume headers. -volt (-v) OR -Voltage Integer Microscope voltage in kV; the default is 300. The program will try to determine this value from the Ctfplotter command file. -cs (-c) OR -SphericalAberration Floating point Spherical aberration (Cs) in mm; the default is 2.7. The pro- gram will try to determine this value from the Ctfplotter command file. -defocus (-def) OR -DefocusInMicrons Floating point The defocus in microns (underfocus positive). The program will try to determine this from the lowest tilt view of the ".defo- cus" file output by Ctfplotter. If that is not possible, the value must be entered. -dcphase (-dcp) OR -PhaseShift Floating point Phase shift in degrees. This entry is needed if you want to account for a phase shift; the program does not take it from the ".defocus" file. -mode (-m) OR -ModeToOutput Integer The storage mode of the output file from either Binvol or Mtffilter; 0 for byte, 1 for 16-bit signed integer, 6 for 16-bit unsigned integer, 2 for 32-bit floating point, or 12 for 16-bit floating point. The default is the mode of the input file. See the Mtffilter man page for some details about mode 12. -setup (-se) OR -SetupChunksIfMemoryError If Mtffilter exits with an error indicating that the memory needed is bigger than the allowed amount, or that memory for the image array could not be allocated, then run Chunksetup to create command files for processing the volume in parallel. -param (-pa) OR -ParameterFile Parameter file Read parameter entries from file -help (-h) OR -usage Print help output -StandardInput Read parameter entries from standard input FILES If the output file already exists a backup of the original is created with the ~ extension. If both reduction and filtering are being done, a temporary file is created with the root name of the output file fol- lowed by ".filttemp" and the extension of the output file. AUTHOR David Mastronarde <mast at colorado dot edu> SEE ALSO Binvol, Mtffilter IMOD 5.2.0 reducefiltvol(1)