Please note that this is an optional step and must be performed only when you have defined ribbons as described in step 2. In summary, you have to perform the same actions mentioned in step 1 but you should use an objective with higher magnification and apply the Global focus strategy under Focus Surface.

  1. You are in step 3/7 Ribbon Imaging.
  1. Under Objective select an objective with a higher magnification than for the overview image, e.g. 10x. The objective should enable you to clearly recognize the structures of interest on your sample.
  2. Under Focus Surface select Global (all Regions/ Ribbons).
  3. Click on Distribute Support Points....
  4. The support points will be distributed automatically over the ribbons. They are displayed as yellow circles with a point in the middle.
  5. If required, you can add further support points by using the Add button below the Distribute Support Points button. Simply click on the image at the position where you would like to add another support point.
  6. Click on Verify Support Points.
  7. Now you can check if each support point is in focus. You will see the overview image in the right image container and the detail image in the left image container. The verification process will start with the first support point which was set. The current support point is marked with a red crosshair. When you activate Show stage position within the image on the Ribbon Definition tab below the Center Screen Area you will see the current position of the stage in the image as a rectangle with a blue dashed frame.
  9. Hold CTRL on your keyboard and use the mouse wheel to adjust the focus for the corresponding support point.

  10. Adjust the focus only by using the mouse wheel. Using the mouse wheel considers the backlash correction. Do not use the focus wheel at the microscope stand.

  11. When the support point is in focus click on Confirm.
  12. The software will automatically move to the next support point.
  13. Repeat the last two steps until you have corrected and verified all support points. At the end of the process you will see the message All points have been verified.
  14. Click on Create Ribbon Image.
  15. The ribbon image will be generated. Note that each ribbon will be displayed as one scene.
  16. Again we recommend clicking on Apply Stitching to remove the offset between the single tile images.
  1. You have successfully acquired the ribbon image and can now continue to the next step by clicking on Next.