Bio-Rad EXQuest Spot Cutter - Manual (Page 532)

Appendix L. Techniques
L-7
indicating that the calibration density decreases while the counts increase (or vice
versa).
To change the orientation of the calstrip so it corresponds to the Calstrip Record
segment activities, use the Horizontal Flip function (under the Image menu). If the
dark segments were on the left, they will now be on the right, and vice versa. Again,
we recommend that you enter your data from highest to lowest activity and display
the calibration strip with the darkest segment on the left.
Box Segments in a Calstrip
Once the calstrip is in the correct orientation, go back to the Edit > Calibrate Gels
submenu and select Box Segment.
Go to the first (leftmost) segment of your calstrip. Sampling boxes must be drawn
beginning with the leftmost segment of the calstrip and moving right.
With Box Segment selected, drag a box on the calstrip. When you release the mouse
button, the area will be sampled.
Note: When defining your box, be sure to avoid sampling regions that are very close to
the borders of the segment or any irregular areas (i.e., areas that are overly dark
or light).
The number of each segment will be displayed in the upper left corner of each
sampling box.
Above the box will be a number between 0.00 and 2.00. This represents the O.D.
value for the segment.
Beneath each box is the number of counts associated with that segment from the
Calstrip Record.
If you make a mistake, simply redraw a new box over the old one. Repeat this
procedure for all the segments of the calstrip.
Note: All segments entered in the calstrip record must be boxed starting from the left.
The leftmost segment corresponds to Segment 1 on the Calstrip Record. Your
calstrips and calstrip data should be designed so that either (1) the darkest
segments are on the left and the count values decrease, or (2) the lightest values
are on the left and the count values increase.