NIH Image Tutorial:
Making a Montage
- Open NIH Image. Use File>Open to view the
images you have selected from the MRI database and
downloaded on your computer.
- 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.
- 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
- To determine the scaling factor of your images
divide their FOV's by 60; this will give you the
"scaling factor". Here are some
So images images from days 6 and 11 which are
80mm need to be scaled by a factor of 1.33.
So images images with an FOV of 70 need to be
scaled by a factor or 1.17.
- Click on the image to be scaled (ex. day 6 whose FOV
is 80mm) to make it active.
- 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.
- Use the rectangular selection tool to select only
the egg in the new window. Select Edit>Copy.
- Click on the original image (ex. day 6) to make it
active. Select Edit>Paste.
- Save the image as a tiff and change the extension to
- Open all of the images you have chosen for
comparision using File>Open.
- Stack them by highlighting Stack>Windows
to Stack in the menu bar.
- Stack>Make Montage to view your chosen
slices side by side. Print out is optional.
- 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.
- Click on the Montage to make it active; then use
File>Save as to save it as either a pict or
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