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Troubles with Tomography » History » Version 1

Amber Herold, 04/23/2010 09:16 AM

1 1 Amber Herold
h1. Troubles with Tomography
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* <link linkend="tomo_intensity">Image appears oversaturated with overflow to negative
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values</link>
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* <link linkend="tomo_intensity">Images acquired in tomography have different
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intensity range from that acquired from other nodes with the same preset and exposure
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time.</link>
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* <link linkend="tomo_large_move">Images move by a large amount between
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tilts.</link>
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h2. Unusual intensity range in the tomography image
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Two possible reasons:
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*  The exposure time has been modified by the tomography node to spread the total dose
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to all images in the tomogram.
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Solution: Reduce the total dose or reduce preset beam intensity. You can find the
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real exposure time from the log panel of the tomography node or through the information
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page of the image in the web image viewers.
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*  The image intensity of images collected by the Tomography class node does not have
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the same meaning as in other images in Leginoon. The flat-field corrected intensity
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values are multiplied by 10 before the image is saved and displayed as MRC image as
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signed 16-bit integer. Therefore any original CCD count of 3276.8 or larger will
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overflow. All other images in Leginon are saved without manipulation as float.
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Therefore, the images obtained through tomography node and another acquisition nodes
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will appear differently in the both the web image viewer and within Leginon.
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Solution: Reduce the total dose.
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Find out what exposure time corresponds to the fractionated dose from your tilt
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angle step and range and total dose and take an image at tomo preset with such an
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exposure in Navigation node. You will need to reduce the total dose if a good fraction
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of the counts are larger than 3200 even though it would not appear to be saturated in
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the float scale without the 10x factor.
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h2. Large movement between tilted images
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Find out how smooth the movement is by comparing the images.
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If transition is smooth and linear:
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*  Magnification too high
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Solution: Lower the magnification or perform <link linkend="Tomo_low_mag_fitting"
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>low-magnification rough model fitting</link>.
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*  Optical axis offset model is too different from the initial guess for the
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non-linear least-square fitting.
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Solution: Perform <link linkend="Tomo_low_mag_fitting">low-magnification rough model
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fitting</link>.
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If transition is not smooth nor linear:
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*  Goniometer behavior unpredictable
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Solution: Service the goniometer.