Appion » TiltPicker

A copy of this page is available at:

  http://emg.nysbc.org/projects/tiltpicker/wiki/TiltPicker_Usage

h2. RUNNING THE PROGRAM:

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To run the program, type:

# ./ApTiltPicker.py -1 image1.mrc -2 image2.mrc -p tiltpicks.spi

'tiltpicks.spi' is optional. I left an example in the tiltpicker folder. You must run the program in the tiltpicker folder. To run the example type:

# ./runPicker.sh

If all works the picker will open up and you'll have two images of ribosomes.

h2. BUG TRACKING:

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 TiltPicker bugs are now tracked via the AMI redmine server, visit

 http://emg.nysbc.org/projects/tiltpicker/

 to file any bugs

h2. YOUTUBE MOVIE:

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I just made a screen capture video on how to use it. It is available is low resolution on youtube:

 http://www.youtube.com/watch?v=cw1osPBY_JM

or you can download the full screen version from the appion.org website:

 http://code.google.com/p/appion/downloads/list

I recommend using VLC to watch it:

 http://www.videolan.org/vlc/

h2. STEP BY STEP TUTORIAL:

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* Start TiltPicker

** Extract TiltPicker, on command line type:	

 tar -zxf tiltpicker-2.0.tgz

** Go to tiltpicker directory, on command line type:

 cd tiltpicker	

** Display TiltPicker help, on command line type:

 ./ApTiltPicker.py -h

** Open TiltPicker, on command line type:	

 ./ApTiltPicker.py -l data/rawu048b.jpg -r data/rawu049b.jpg -t 50.0	

* Initialize alignment:

** Maximize window

** Set correct zoom for images

** Center the middle bar

** Pick 5 partcle pairs

** Save picks to a file

** Hit the apply button, circles will not match crosses

** Find theta (tilt angle)

** Optimize inplane rotations

** Optimize all parameters

** Mask region to only look at overlapping regions, overlap is shown on status bar

** Hit Auto optimize, it performs find theta, optimize angles and mask in a single click

** If you look at the Terminal it will tell you how many optimization rounds were performed

** Use transfer picks button, this works but is based on the alignment parameters

* Auto pick particles

** Measure particle size using ruler tool, zoom in to get best value

** Run DoG picker with wrong contrast

** Reject picks

** Run DoG picker with correct contrast

** Show terminal

** Accept picks

** Look at DoG map images

* Clean up particle picks	

** Use clear bad picks button	

** Use polygon tool

** Auto optimize

** Use cutoff button to remove picks below a threshold

** Add more picks

** Save and quit