NIH Image Tutorial:
Making a Montage

1. Open NIH Image. Use File>Open to view the images you have selected from the MRI database and downloaded on your computer.

2. Using the control parameters you have set, choose the "best image" from each of the five days. The image files must be named MRI01.jpg, MRI02.jpg....MRI10.jpg, etc. for NIH Image to stack them in order.

3. All images used to compare the area of the eye should be the same scale or "resolution" in pixels/mm. If you have chosen images with differing field of views (FOV), you will need to scale some of them. (You recorded the FOV earlier when you downloaded the image from the MRI Database.) For example: The MRI instrument for days 7-10 is "zoomed in" to a 60mm field of view and the FOV for days 6 and 11 is 80mm. The eye is easier to see on the "zoomed in" images. Images with an FOV of 60mm work well for this lesson. To scale images to a uniform resolution which will match those with an FOV of 60mm, use the following steps:

1. To determine the scaling factor of your images divide their FOV's by 60; this will give you the "scaling factor". Here are some examples:

 80.0/60.0=1.33 So images images from days 6 and 11 which are 80mm need to be scaled by a factor of 1.33. 70.0/60.0=1.17 So images images with an FOV of 70 need to be scaled by a factor or 1.17.

2. Click on the image to be scaled (ex. day 6 whose FOV is 80mm) to make it active.

3. Edit>Scale and Rotate will bring up the following pop-up box:

Fill in both Horizontal Scale and Vertical Scale boxes with the "scaling factor" determined in step a (1.33). The new larger image will appear in a new window.

4. Use the rectangular selection tool to select only the egg in the new window. Select Edit>Copy.

5. Click on the original image (ex. day 6) to make it active. Select Edit>Paste.

6. Save the image as a tiff and change the extension to .tif.

4. Open all of the images you have chosen for comparision using File>Open.

5. Stack them by highlighting Stack>Windows to Stack in the menu bar.

6. Stack>Make Montage to view your chosen slices side by side. Print out is optional.

7. Click on the Image stack to make it active, then use File>Save as to save it as either a stack of picts or tiffs.

8. Click on the Montage to make it active; then use File>Save as to save it as either a pict or tiff.

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