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Creating a new lens based on a calibration grid slide

Procedure

  1. Acquire an image of the MBF calibration grid slide using the same objective that you plan to use in your study.

    If you are using multiple objectives, acquire an image of the calibration grid slide with each objective.

    • 10X and lower: Acquire an image of the large grid (250 µm).
    • 20X and above: Acquire an image of the small grid (25 µm). Immersion media (oil, water, etc.) are not required for this step.

  2. Load the grid(s) image into the program. The Image Scaling dialog box opens .

    If Current lens is selected under Source Of Scaling, check the Override X and Y scaling box. This will enable you to define an appropriate scale factor when you create the new lens (step 5).

  3. Open the Image Organizer.
  4. Make sure only the relevant grid image is visible:

    For example, if you are calibrating a 5X objective, the icon in the Image Organizer should only appear next to the 5X grid image.

  5. Click the File>Calibration>New lens button.
    • If the image is not displayed at its original magnification, it is automatically adjusted to display at a 1:1 resolution.
    • Follow the instructions for Creating a New Lens.

    If you are calibrating from an image with a scale bar instead of the MBF calibration grid slide, check the Force Square checkbox.

  6. If you need to define multiple lenses from calibration grid images, hide the image for the lens that has just been defined and repeat this process for the other lenses.
  7. Use the measurements functions (Trace>Measure line drop-down) to verify that the calibration is correct.

 

See Calibrating your System