Bio-Rad Components for Older Model Spot Cutter - Manual (Page 16)

Specify the microtiter plate size you are using by clicking on 96 wells or 384 wells.
Specify the diameter of the cutting tip by selecting 1.0 mm or 1.5 mm. Note that the cut
request circles that you make on the image reflect the actual size of the cutting tip, and will
change based on the tip size selection.
Specify the order in which you want to the load the wells on the microplate by selecting A1,
A2, A3, … or A1, B1, C1, …
Crop Left Margin repositions the image in the Basic Excision Tool window to the left. Enter
a value (in screen pixels) in the field to reposition the image. Note that you must click on
Done to close the settings dialog and then acquire a new image for this change to take effect.
Well A1 position
The fields and buttons in this section are used to calibrate the position of the microplate for
proper loading of samples. The number fields contain the X and Y coordinates (in millimeters)
of well A1 on a plate in the spot cutter plate holder. Once you have set the position of this well,
the cutter robot will automatically "know" the positions of the remaining wells.
To set the well A1 position, position your cursor over the number in the X or Y coordinate field
and press Enter on your keyboard twice. The tip will move to that position without attempting
to lower into the well. If the tip is not centered directly over the well A1 on the microplate, you
will need to adjust the numbers in each field.
You can either type a new value directly in each field, or (for small adjustments) use the arrow
buttons to adjust the tip by fractions of a millimeter. The cutting tip will move as you either
enter new numbers or click on the buttons.
Note that the arrow buttons are oriented as if you were facing the front of the spot cutter; also,
if you are facing the cutter, higher numbers in the first field will move the cutting tip to the
right, while higher numbers in the second field will move the cutting tip toward the back of
the instrument.
When you are satisfied with the position of the cutting tip, click on Test to lower the tip into
the well. The tip should be centered inside the well.
Park returns to the tip to its home position.
Wash Station 1 position
Calibrate the position of Wash Station 1 on the cutting platform by following the steps
described above for Well A1. Once you have set the position of station 1, the remaining wash
station positions are automatically calibrated.
Spot Cutter Calibration
Before you can begin excising spots, you must calibrate the spot cutter. This procedure requires
Spot Cutter Gel Sheets and a coomassie blue stained PVDF membrane.
Note: Before calibration, you should focus the cutter camera and align the wash station
position. You should make sure the spot cutter camera image is properly centered using the
Crop Left Margin setting.
Click on the Calibrate Cutter button. A series of dialog boxes will guide you through the
calibration procedure. At any point in the procedure you can move back to repeat a step.
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