This topic describes how to calculate the alignment parameters for the DeltaVision OMX.
The following figure shows a target image before calculating the alignment parameters (left image) and after (right image).
IMPORTANT Since system alignment parameters are different for each dichroic drawer, this procedure must be completed for each drawer used with the system.
• Crosshair objective (Part Number 57-750795-005)
Ensure that the installed dichroic drawer has been aligned, as described in the DeltaVision OMX User Guide.
To calculate alignment parameters for the DeltaVision OMX:
1. Carefully remove the imaging objective from the DeltaVision OMX and install the crosshair objective. (Refer to “Removing and Replacing the OMX Objective” in the DeltaVision OMX User Guide.)
2. Ensure the transmitted light arm is positioned over the objective.
3. Using the DeltaVision OMX software, acquire a transmitted light image (DIC) of the crosshair. Ensure this image has the following characteristics:
Consists of a single Z section
Consists of a single time point (no time-lapse)
Includes all available channels
4. In the softWoRx main menu bar, click Process | OMX Alignment Parameters.
5. Specify the target image acquired in Step 3. Choose one of the following:
Click Calibration Image and select a file from the displayed list. If necessary, use the Directory field and/or navigation icons in the Select File dialog box to navigate to the directory containing the desired file.
OR
Drag-and-drop a file from the Konqueror file manager into the Calibration Image field.
OR
Type the path and file name directly into the Calibration Image field.
The OMX Alignment Parameters dialog box reappears, this time containing field data.
6. Click Drawer to Calibrate and select the appropriate drawer from the drop-down list.
7. Click Do Alignment Calculation. After a few seconds, the aligned target image appears in a new image window.
NOTE Z shifts may also be stored with the calibration data, though they will not affect the aligned target image.
8. When the alignment is complete, click Save Results.