Before you can start to acquire S&F images you first have to calibrate the correlative holder. Note that holder calibration must be performed on both systems (LM and SEM). During calibration the exact positions of the three small fiducial L-shaped markers must be detected (or set manually). For each marker you can decide if you want to use the automatic marker detection or if you want to manually set the marker position, e.g. if the auto detection failed. Since the auto detection usually works fine, we recommend always using it.

1. You have inserted the holder to be calibrated on the stage (example LM system).

1. Open the S&F Holder Calibration workbench.
2. In the Calibration Settings select the corresponding holder from the drop-down list.
   Make sure that the Holder Orientation setting within the software is the same like on the stage. If this is not the case, change the orientation in the software. We also recommend to activate Automatic movement to next marker, so the process will be more fluent.
3. Click Start on top of the workbench.
   
4. A dialog will appear to choose whether you want to perform automatic or manual marker detection. We recommend to use the automatic marker detection which is performed as follows:
4. Move the stage in the area of the first marker and click Auto . Make sure that the Live image is in focus and the marker is visible.
   
5. The software will now try to detect the first marker on the inserted holder. The marker is detected correctly, when the red crosshair matches the corner of the smaller L-shaped marker exactly. After the detection a confirmation dialog will appear.
   
5. Click Yes, if the marker was detected correctly.
   If the marker was not detected correctly, click No and select Manual mode. Set the marker manually by left clicking with mouse on the outer corner of the small L shaped marker. The position will be saved and you can continue to the next marker.
   Repeat the last two steps for the next 2 markers. The dialog box will always show you which marker is to be detected.
   
6. If all three markers were detected correctly, the calibration is completed.
6. Click Finish Calibration to close the wizard.
   

1. In the Calibration Settings tool you now will see the position data (X/Y) of the first marker. If the holder was calibrated once, you can always move the stage easily and fast to the first marker position by only clicking 1.Marker on top of the workbench.
   
2. You have calibrated the holder successfully. You can now acquire images of your sample. Note that the calibration process must be performed again, after the holder is transferred (shuttled) to the other system. Otherwise the sample positions (ROIs/POIs) cannot be found automatically.
3. After you have completed the "default" calibration procedure once, you can perform an automated calibration, e.g. if you have changed the sample on the calibrated holder. To learn more about the auto-calibration, read the chapter Calibrating the S&F Holder Automatically.