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Load the images you would like to examine as a group (see Load Groups of Images Tech Note 12).

1. Open the image you would like to use for drawing your background ROI. 2. Select Average Bkg ROI from the Type dropdown menu under ROI Tools.

3. Draw your background ROI in an area of the animal representative of average background levels.

4. Double click on the BKG 1 ROI and the ROI Properties window will open. 5. Check Use as BKG for future ROIs in, then select Entire Sequence. Every

subsequent measurement ROI drawn on images in that sequence will automatically subtract the average background as calculated from the BKG 1 ROI.

Subtracting Background ROI from a Sequence

Tech Note 5b

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6. Switch back to Measurement ROI in the Type dropdown menu in ROI Tools

and draw your measurement ROIs. You will notice that each measurement ROI is linked by the presence of (BKG 1) in the measurement window and in the ROI Measurements table which denotes the specific background ROI has been subtracted correctly.

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Tech Note 5b www.CaliperLS.com