Bio-Rad Components for Older Model Spot Cutter - Manual (Page 13)

Recutting Spots
If the cutting tip has not picked up a spot, click on the Recuts button below the image window.
In the pop-up box, enter the numbers of your recut requests (microplate number, well
number), separated by semicolons. Click on OK to close the recut request dialog, then click
on Begin to perform the recuts.
Acquiring the Image
Before acquiring an image using the spot cutter camera, select the Light On option if you are
imaging a gel, or Light Off if you are imaging a PVDF membrane. (The platform light should
be off for PVDFs, because the ambient light will produce a better image.)
Click on Acquire Image. The camera will take a single image of the gel or membrane on the
platform, and the image will appear in the window.
Zoom and Transform
On the right hand side of the screen are the zoom controls. The first icon on the top is used to
return the image to full view. The second icon down is used to magnify a region specified by
click-dragging a box around the region. The third icon down (the drag hand) grabs the image
to move to different locations within the image. The fourth icon down zooms in centered on
the placement of the icon, the fifth zooms out in the same way. The transform icon is the last
in the column of icons, and is used to adjust the brightness and contrast of the image.
Specifying Cuts
In this step, you will click on spots in the image to identify which spots to cut.
When clicking on spots in the image, select the Snap to Peak checkbox to automatically
center the cut circle on the spot peak (calculated from the spot intensity in the image). You can
also use the Zoom Box tool included in the Basic Excision Tool to magnify each spot.
To make a single cut in a spot, make sure the Multiple Cuts/Spot checkbox is unchecked,
then click on the Add Cut button {insert add cut.gif} and click on the spot in the image that
you want to cut.
If this is the first cut circle marked on the image, the Enter Plate Information dialog box will
open, and you will be prompted to enter a name for the microplate and identify the first well
you want to load. If you specify a starting well other than the default of A1, the skipped wells
will be noted in the dialog box. Click on Done to make your selections.
A green circle will appear on the image, indicating the spot to be cut. Note that the circle is
the size of the actual cutting tip (1.0 mm or 1.5 mm, as specified in the Spot Cutter Settings
dialog box).
Continue clicking on spots in the image with Add Cut selected to make additional cuts. The
number of cuts will appear in the Cuts Selected field.
If you specify more cuts than there are wells in the microplate, you will be prompted to name
a new plate and starting well for subsequent cuts.
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