Making Composite Images

  1. Open Image J from the Applications Menu in the Finder Window
  2. Open all 3 of your images by dragging their icons to the ImageJ toolbar.
  3. Open the Image menu, and select color>merge
  4. Put your rhodamine (phalloidin) image in the red channel, your fluorescein (FITC) image in the green channel, and your DAPI image in the blue channel. Leave the other channels blank.
  5. Save the composite image both as a TIFF and a JPEG with a reasonable file name, but don't discard the originals. The TIFF will work better if you decide to do further analysis in ImageJ, but the JPEG may work better for your lab report.

If you took the three images without moving the specimen, this image should show you the relative distributions of dsDNA, actin, and the cellular protein your chosen antibody bound to in the cells.