- Define a new project space with the name "KLH".
- Load micrograph "KLH_02_01.mrc".
- Quality inspection: intensity histogram seems okay.
- Quality inspection: activate SPEC
button, inspect spectra of selected regions.
- Filter the image to reduce noise and enhance contrast. In
Image
Pre-processing box, set Low-pass
to 140, and High-pass
to 1000. Click
the Filter
button.
- The templates will come from the micrograph
directly.
Click the Filament
button at the bottom of the window. A rectangular
monitor will show up in the panel.
- Set particle diameter (Size)
to 800 in the Filament
Parameters box.
- Identify a clean top-view, draw a box around the particle
(press CTRL, left-click and drag). Set the scaling of the monitor to 2
or 3 in
order to display the full particle. Save the image (Save
button) as "topView.mrc" (see
example).
- Identify a clean side-view, draw a box around the particle,
and save the image as "sideView.mrc" (see example).
- Go to Menu
: Function : Image Stack Editor, click New
and load "topView.mrc". Then, click Add
to append "sideView.mrc". Crop both
images to an appropriate size (200-pixels in the example) using the Crop
function in the Image
Processing box. Click Save
to save the templates as "klhTemp.mrc" (see example).
- Load template image "klhTemp.mrc" (Menu
: Data : Template : Load
MRC).
- Erase the default template masking by clicking Clear
in Manual
Masking box.
- Set Frame
to 1 in Manual
Masking box, then click Edit.
The top-view is
displayed. Activate the circle-drawing button on the toolbar (mouse
click or press F3 key). Position the circle around the top-view at a
proper radius (Left-click-drag for shifting, Right-click-drag for
radius resizing). Deactivate the circle-drawing button, a circular mask
will be drawn. Release the Edit
button.
- Set Frame
to 2 in Manual
Masking box, then click Edit.
The side-view is
displayed. Activate the line-drawing button on the tool bar (mouse
click or press F4 key). Set the brush-size to 16 and draw a rectangular
mask around the particle. Deactivate the line-drawing button, release
the Edit
button (see example).
- Micrograph screening can now start following the normal
procedure.
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