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Add Scale Bar in ImageJ: Step-by-Step Guide

This document provides step-by-step instructions for adding a scale bar to images using ImageJ software. It outlines the process of calibrating the scale by measuring a stage micrometer at different magnifications and setting the scale in ImageJ. Finally, it explains how to add and customize the scale bar on the desired image before saving it.

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522 views2 pages

Add Scale Bar in ImageJ: Step-by-Step Guide

This document provides step-by-step instructions for adding a scale bar to images using ImageJ software. It outlines the process of calibrating the scale by measuring a stage micrometer at different magnifications and setting the scale in ImageJ. Finally, it explains how to add and customize the scale bar on the desired image before saving it.

Uploaded by

oemoncur22
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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Download as DOCX, PDF, TXT or read online on Scribd

ImageJ Software: Instructions to Add a Scale Bar

Image the stage micrometer using the ocular camera for each of the scope objectives (10x, 40x
and 100x with oil). This is going to use the “ruler” in order to make a pixel to distance
conversion that will ultimately allow you to add a scale bar to all of your cell biology images.

1. Download the ImageJ software to your computer (choose appropriate operating system):
https://imagej.net/ij/download.html

2. Open ImageJ. When you open the program, you will get a window similar to this (some
buttons depend on the version):

We will be using the “line selection tools” for most of our work. Note: There are a lot of tools
within this software package that may be beneficial in annotating your figures. ImageJ is a
common software that scientists use.

3. Open your image of the micrometer (this is so you can set the calibration scale). To open your
images: Choose File > Open, navigate to your folder that contains the image and select it.

The image to the left is of the whole scale on the micrometer


slide. It shows 2mm total, with 0.01 mm subdivisions
between 0-1.0, and then 0.01 mm subdivisions in the very
small hashes between 0-0.1. The circle represents what part
you will actually see under our microscopes with the 10x
objective.

4. YOU MUST tell the software the scale in order to have


your scale bar be correct. Select the straight line selection
tool. Draw a straight line that defines a known distance on
your micrometer image. To measure accurately, enlarge your image and draw a line from the
outside of one black line to the inside of the next line (as shown below)

5. Choose the Analyze > Set Scale. The following dialog box will open.

The distance in pixels will be filled in for you based on the length of the line you drew in step 4.
Now you must fill in the Known Distance box based on the ruler length at that magnification (so
that could be 0.1 or 0.01 depending on what distance you measured. Then define the unit label in
the Unit of Length box (most of the time this will be mm). If you check the Global box, every
image you open in this ImageJ session will have this scaling. Click OK. **remember that you
need to change this between 10x images, 40x images and 100x images.**

6. Now open the image you want to add a scale bar to. Choose the Analyze > Tools > Scale Bar.
You can adjust the size, color, and placement of your scale bar. Design and place a scale bar that
will best work on your image. Once you are finished click OK and then save your image.

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