quickly scan through a file series to check if the automatically determined defocus values make sense.
Typical use:
1) On a set of micrographs, generate 2D power spectra with the batch file power.spi.
2) Estimate defocus with the TF ED command via the defocus.spi batch file. This outputs a set of doc files with various curves, as well as a doc file containing the defocus estimates.
- 3) Run ctfmatch.
- - Set the correct parameters,
- read in the set of curve files generated by TF ED,
- read in the defocus doc file,
- select a file to display. Adjust the model parameters if necessary.
- You can save the new defocus values to a file, if you want.
Command line usage
Usage: ctfmatch [tfed_file] [-arg value]
or for multiple files: ctfmatch -tfed file_pattern*
where [tfed_file] is an output doc file from the TF ED command with 4 columns:
[spatial frequency, background, subtracted data, envelope ]
The remaining arguments are (-keyword value) pairs, where the keyword is denoted with a minus sign.
- The following keywords are supported:
- -cs (spherical aberration)
-kev (electron energy)
-pixsize (A/pixel)
-src (source size)
-spread (defocus spread)
-acr (amplitude contrast ratio)
-gep (Gaussian envelope parameter)
-tfed file_pattern*
Examples:
ctfmatch -cs 2.20 -kev 200 -pixsize 2.82
ctfmatch -tfed ctf00*.dat
ctfmatch -help (prints help message)
Controls
There are a number of slider controls that interactively alter parameters in the displayed curves.
- Model sliders
- As in ctfdemo, values for defocus, source size, defocus spread, and the Gaussian envelope may be adjusted with the slider controls. Other parameters (pixel size, electron energy, spherical aberration, and amplitude contrast raio) are set in the Options :: Parameters menu. These values only affect the model data (shown in the white curve).
- Graph sliders
- Because of the great dynamic range of the data, the ranges of the graph can be adjusted:
- ymax : adjusts the maximum value on the vertical axis
- model height : adjusts the height of the model data
- xmin : adjusts the left hand side of the graph
- reset ymax button : when xmin of the graph is changed, the data can be rescaled accordingly with this button.
- Show Menu
- The Show menu lets you turn the various curves on or off. A curve will be displayed if its box is red (or a checkmark on some systems). As curves are toggled on and off, the maximum height (ymax) of the graph is automatically adjusted to use the entire area for display.
Options
- Parameters
- lets the user set values for pixel size, electron energy, spherical aberration, and amplitude contrast ratio.
- Squared data
- Because the procedure file power_p1.bat takes the square root of the spectrum, ctfmatch is automatically squares the TF ED data it reads in. This is to ensure that the model parameters are in the same units as the data. Thus by default, the Squared data box is checked. Unchecking this box displays the square root of all the curves (i.e., the unsquared data).
- Grid
- toggles a grid on and off.
- Use empirical envelope
- Forces the model to fit under the empirically derived envelope (the blue curve). When this option is in effect, various model parameters affecting the envelope, such as source size and defocus spread, are no longer used to compute the displayed model curve.
References
For details about the TF ED command, see the manual page, and
Z. Huang, P.R. Baldwin, and P. A. Penczek. Automated determination of parameters describing power spectra of micrograph images in electron microscopy J. Struct. Biology Vol. 144, Issues 1-2, October 2003, pp. 79-94.
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