Living Image 3.1

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6. Image Math 102 To subtract tissue autofluorescence: 1. Load the image sequence that includes the primary and background fluorescent images. 2. Open either the primary or background image and: a. Optimize the image display using the color scale Min and Max sliders in the Image Adjust tools. b. Draw a measurement ROI on an area of the animal that represents background signal (area where no fluorophore signal is present). Note: You only need to draw the ROI on one of the images. The software copies the ROI to the other image. 3. Select Tools → Image Math for xx.SEQ on the menu bar.
Living Image ® Software User’s Manual 4. In the Image Math window that appears, select the primary image in box A. Select the background image in box B. Note: For more details on items in the Image Math window, see Table 6.1, page 100. 5. Select the math function 'A-B*k' in the Result drop-down list. 6. Click and select the ROI (created in step 2) from the drop-down list. - The background-corrected signal is displayed. 7. To view the mathematical result (overlay mode) in a separate image window, click Display Result For Measuring. Note: If necessary, use the Color Scale Min and Max sliders in the Image Adjust tools to adjust the image display. To save the new image: 1. Click the Save button . Alternately, select File → Save on the menu bar. 2. In the dialog box that appears, select a directory, and click Save. - A folder of data is saved to the selected location (AnalyzedClickInfo.txt, ClickInfo.txt, luminescent and photographic TIF images). To export the new image to a graphic file: 1. Click the Export button . 2. In the dialog box that appears, select a directory, enter a file name, and select the file type from the Save as type drop-down list. 3. Click Save. 103
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Living Image 3.1

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