Setting Up Ratio Imaging

softWoRx provides a ratio imaging acquisition function that allows you to view a graphic representation of the ratio of two channels as the images are being collected. In addition, a ratio graph displays the mean value (of an area in the middle of the image) vs. time. Both the ratio image and the ratio graph are for monitoring purposes only. The ratio imaging experiment results in a two-channel time-lapse image.

To set up a ratio imaging experiment:

  1. From the softWoRx main menu, select File | Acquire (Resolve3D) to open the Resolve3D window.

  2. On the Resolve3D window, click the Experiment Icon_Experiment.jpg icon to open the Design/Run Experiment window.

  3. Select the Sectioning tab and deactivate the Z Sectioning check box.

  4. Select the Channels tab and specify the two channels you want to use for this experiment.

  5. Select the Time-lapse tab and specify the time-lapse and total time for this experiment.

  6. Select the Actions tab and select Ratio Image as the action for this experiment. The Time Points specification will default to all and the When control will be After Imaging.

  7. Select the Run tab and enter the image file name and a title for the ratio image. You can also enter text into the Add note to log field to include the text in your image log file.

  8. Click Start Scan Run_icon_sm.JPG to begin the imaging process. The ratio imaging process will occur in a separate Image viewer. (The square outline in the center of the image represents the mean value of the image data.)

When the ratio imaging experiment is complete, a two-channel time-lapse image is displayed along with a ratio graph showing the mean value (the outlined area in the middle of the image) vs. time. The square outline represents the portion of the image used to calculate the mean for a ratio graph.