Bio-Rad Components for Older Model Spot Cutter - Manual (Page 21)

7. You should be able to see "holes" in the spots in the image within the overlay target circles.
However, there is a + or - 6% to 10% accuracy spec (depending on tip size), so an edge of
the "hole" may be outside the circle and still be within specification.
Note:
The "snap to peak" and "gaussian fit" targeting features center the cuts on the densest
portion of the spot, not necessarily in the visual center of the spot. If the target circles and
cuts are off center, this does not necessarily mean that the targeting and cuts are off the
intended cut.
8. When you have viewed the cuts, click on the “Begin/Resume” button. This will close the new
image window with the overlays, and will prompt for loading the microtiter plate. Click “OK“
when you are ready to resume the cut run.
9. The cut run will resume on the next spot to be cut using the original cut targets.
10. You can repeat the "Confirm cuts" operation as many times as you want during the cut run.
The "confirm cuts" is an option in fluorescent gel cutting. Once you are familiar with targeting and
cutting fluorescent gels, you may or may not continue to use this function.
Section 5
Fluorescent Operation in Integrated Excision Tool
The PDQuest version 6.2.1 is very similar to version 6.2.0 in operating the spot cutter. The main
differences are some new function buttons and workflow items based on using the fluorescent
enclosure. For a more complete description of the Integated Excision Tool using PDQuest image
analysis, see the PDQuest manual. The following sections describe the workflow of the
Fluorescent Enclosure items that differ from the white light operation for the Integrated Excision
Tool.
5.1 Calibration and the UV Light Option
During calibration of the spot cutter, the bandpass filter must be installed if UV/fluorescent imaging
is going to be used for spot cutting. In order to record that the bandpass filter has been installed
for calibration, the first question that comes up in the software during calibration is to confirm that
the lens bandpass filter has been either "Installed" or "Not Installed". Once the system has been
calibrated with the "Filter is Installed" message answered, the "UV on (Fluor)" light option is active.
If the question is answered "Filter is not installed", then the "UV on (Fluor)" light option will not be
active.
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