Quantification of Vacuolar Colocalization

Software

• ImageJ - available at http://rsbweb.nih.gov/ij/

Method

1. You Need A Merged High Resolution Image
2. Open File in ImageJ
3. Go To Image -> Color -> Channels Tool
4. From the dropdown menu select Color then click OK
5. Zoom in to see the cell
6. Draw a Line using the Line tool. The line should start inside the cell, and cross the vacuole twice. Try to pick a representative cross section of the vacuole
7. Select Analyse -> Plot Profile (or hit the shortcut CTRL-K)
8. On the popup window select copy and paste the data into an excel document
9. Move the slide bar on the bottom of the image to the green channel
10. Select Analyse -> Plot Profile (or hit the shortcut CTRL-K)
11. On the popup window select copy and paste the data into an excel document