— Sean Delica 2021/02/10 14:48

Note: Magnification of the wormscope needs to be calibrated using a micrometer, if that has not been done yet.

1. Go to amscope application, which has the data for your magnification specs.
2. Click options → magnification → click on the setting you used to take photos that you wish to put the scale bar on, then press edit. Copy the resolution indicated on the edit tab, which will likely be in pixels/meter.
3. Calculate that value in pixels/meter and change it to pixels/micron. We will be using that value in imageJ for calibrating the scale bar. (1 meter = 1,000,000 microns)
4. Open imageJ. Click analyze → set scale → enter the calculated values from step 3. Keep pixel aspect ratio to 1. Check mark “Global”. Done properly, the window should show the same values you calculated.
5. Make a folder for all the photos, either cropped with a specific pixel dimension or uncropped, that you want a scale bar inserted. It would be preferable for those photos to have the same resolution.
6. A window with all your photos should show. Now, click analyze → tools → scale bar. You can adjust these settings to what you see fit. The window showing all the photos will preview what the scale bar would look like according to the input values.
7. To save your photos, file → save as → image sequence. Change the format to your liking. Check mark “use slice labels as file names” to keep the same names of each photo.