Bio-Rad Quantity One 1-D Analysis Software - Manual (Page 122)

Chapter 5. Bands
5-5
The Detect Bands dialog has a short format and an expanded format; toggle between
them by clicking the toggle box in the lower right corner.
To change a parameter in the dialog, you can either type in a new value or use the
arrows to increase or decrease the setting by 10 percent. Experiment with different
settings to find those best suited to your images.
5.2.a Detection Parameters
Lanes to Detect
If the intensities of the bands vary from lane to lane, you may need to use different
detection parameters on different lanes. Specify whether you want to use the detection
parameters for all the lanes or a single lane by selecting All or One next to the Lanes
prompt. If you choose One, type the lane number in the field next to the One button.
When to Detect
If you select Auto next to the Detect prompt, band detection will occur immediately
each time you change a detection parameter. You will not need to click on the Detect
button located at the bottom of the form.
To change more than one parameter before detecting, choose Manual. With the
Manual option, you can change parameter settings first, and then apply them by
clicking the Detect button.
Normalization
Normalization is a way to compensate for differences in intensity between lanes. It
does not normalize for band quantitation.
The intensity of each lane is determined by the darkest band in that lane. For example,
suppose that in all but one of the lanes the darkest band has an intensity of 50,000
counts. In the one light lane, the darkest band is only 25,000 counts. With
normalization, band detection will be twice as sensitive when processing the light
lane, improving the detection of faint bands.