Bio-Rad EXQuest Spot Cutter - Manual (Page 139)

PDQuest User Guide
4-18
Note: It is also possible to add spots on Raw 2-D scans when the Filtered and the
Gaussian images are loaded.
Fig. 4-11. Removing incorrectly identified spots in a cluster.
4.6 Spot Boundary Tools
If a spot is saturated, irregularly shaped, or otherwise of poor quality, then Gaussian
modeling will be unable to accurately determine quantity. In these cases, you can
manually define the spot in the Filtered or Raw 2-D image using the spot boundary
tools.
Note: For most spots, Gaussian modeling results in more accurate quantitation than
defining the spot boundary.
When you define a spot boundary, the region inside the boundary is identified as the
spot and is added to the Gaussian image. Spot quantity is calculated by summing the
intensities of the pixels inside the defined boundary. You can define the boundary of
any spot in a Filtered or Raw 2-D image.
Solution: Use Remove Spot, then Result: Correct number of spots in the
Problem: Too many spot centers
identified in the Filtered image. Result: Too many spots appear in
Gaussian image.
Add Spot to reposition spot centers. Gaussian image.