When your software is correctly configured, all images that you acquire with your software are calibrated automatically. This magnification doesn't, however, correspond with the actual magnification of the acquired images. The actual magnification depends on the camera's position. The magnification and therefore the image calibration will vary depending on whether the camera is bottom-mounted or side-mounted.
Use the Magnification calibration process to acquire correctly calibrated images. Correctly calibrated images are a necessary precondition for being able to measure, for example, distances and areas on your images.
Starting the software wizard
To start the software wizard for this calibration process, select the Magnification calibration process, in the Acquire > Calibrations dialog box. Then click the Calibrate... button.
Description of dialog box
The Calibrate Magnifications dialog box is divided into different areas. Click on one of the areas in the dialog box shown schematically above to jump to the corresponding help topic.
(1) Chart and table showing the current calibration data
(2) Preparing the calibration process
(3) Performing the calibration process
See also
(1) Chart and table showing the current calibration data
Check the current calibration data in the Calibrated Magnifications group.
In the table, you can find all the calibration processes that have already been carried out and their corresponding magnifications. The magnification and the resulting X and Y resolution is specified in units of [pixels/nm].
The chart shows the reciprocal pixel size plotted against the magnification. The points should all be roughly on one line.
Deleting calibration data
Select the entry in the table containing invalid calibration data, and click the Delete button.
When you carry out a calibration process for a magnification that already has calibration data, the new calibration data replaces the existing calibration data.
How many magnifications have to be calibrated?
Repeat the calibration process for at least three different magnifications.
You don't have to carry out the calibration process for all of your microscope's magnifications. When you set a magnification that's not in the list, the calibration data is extrapolated from the available calibration data.
(2) Preparing the calibration process
The Preparation group describes the calibration process flow.
Selecting the calibration standard
To be able to carry out the calibration process, you need a calibration standard that has structures of a know length. You can use the Carbon Black calibration standard or a commercially available cross grating standard, for example.
Select the calibration standard that you are using from the list.
Editing calibration standards
When you are using a calibration standard that isn't on the list yet, you can define an additional entry for the list.
1.In the Preparation group, select the Edit Calibration Standard... entry from the list.
2.The Edit Calibration Standard dialog box opens.
2.In the Edit Calibration Standard dialog box, click the Add... button.
•Another Edit Calibration Standard dialog box opens.
3.Enter the name of your calibration standard in the Name field.
In the Reference length field, you enter the length of the structure that you want to use for the XY calibration.
During the calibration, you can easily use multiples of the reference length to enable you to use a calibration standard for a large magnification range.
4.Close the two Edit Calibration Standard dialog boxes with OK.
•The new calibration standard will then be displayed in the list.
You can delete self-defined calibration standards from the list or change the specified reference length at any time.
1.In the Preparation group, select the Edit Calibration Standard... entry from the list.
2.In the Edit Calibration Standard dialog box, select the calibration standard you want to edit or delete.
3.Click the Delete or Edit... button.
•The predefined calibration standards can't be deleted or edited.
Calibrating on live-image
Select the Calibrate on live image check box. Now, you can carry out the whole calibration process in the live mode.
If you don't want to define the reference length in the live-image because the live-image is moving too much, clear the Calibrate on live image check box. Now you can focus the sample, select the exposure time and select the microscope magnification in the live mode. Then click the OK button in the Calibration Note message box to acquire the image to be used for the definition of the reference length.
Calibrating the magnification using a Fourier transform
Select the Calibrate via Fourier Transform check box to calibrate the magnification using a Fourier transform. Calibrate using a Fourier transform when you are working with very high magnifications. You can find a step-by-step instruction here.
(3) Performing the calibration process
Click the Set Calibration Distance... button to start the calibration process. You can find a step-by-step instruction here.
Click the Finish button to complete the calibration process. For this to work, you have to have carried out the calibration process for at least three different magnifications.
If fewer calibration data are available, you are able to leave the Calibrate Magnifications dialog box, but the calibration process remains incomplete. Then the dialog box will contain only the Finish button and not the OK button. The acquired calibration data remain unchanged.
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