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Camera/stage alignment

1. Verify that the camera is oriented in the proper direction: What appears through the eyepieces should be identical in orientation to what appears on the screen.

   Example with the MBF calibration grid slide

   If the small grid is to the left of the large grid when looking through the eyepieces, then it should appear the same way on the computer screen. If it appears backwards, you need to rotate the camera 180°.

   • To adjust the camera orientation, loosen the set screw and rotate the camera. Tighten the set screw when finished.
2. Place a reference point on the screen (anywhere is fine).
3. Click the Display Grid icon ; a white grid appears on the screen, displaying horizontal and vertical reference lines.
   • For this step, refer to this new virtual white grid on the computer screen (instead of the black grid on the grid slide).
   • To adjust the grid size, use the Options>Display Preference>Grid tab. Adjust the X and Y Grid Spacing measurements as needed)
4. For a preliminary alignment, at a low objective (2X or 5X), locate the large grid on the slide (250 µm) and align it closely to the grid on the screen .
   • Right-click and select Joy Free to control the slide movements with the joystick.
   What do I do if the two grids don't line up?

   Adjust the camera:

   1. Loosen the set screw (sometimes located at the bottom of the C-mount).
   2. Slowly, and only slightly, turn the camera in either direction until the two grids line up on the screen (make sure not to make large rotations with the camera or it may become oriented backwards).
   3. Tighten the set screw to set it into place; this provides a rough starting point for camera alignment. Aligning the grids is NOT sufficient for proper calibration.
5. Continue in Joy Free mode at a higher objective (10X or 20X).
   • Navigate around the grid slide to find a dust particle easy to identify (e.g., particle with a defining mark or protrusion) .
6. Move the dust particle to the top of a white grid line on one side of the screen.
   • Using the joystick, move the particle across the screen to the other side on the same grid line. With a perfectly aligned camera, the dust particle will remain lined up with the grid line.
     • If it has moved above or below the line, adjust the camera to correct for the movement: Loosen the set screw and adjust the camera before tightening the screw again.
7. Continue moving the dust particle across the screen and adjusting for correction if needed.
   • Once the dust remains constant across the screen, the camera is aligned.
8. Move to a higher objective and repeat steps 5–7 for a more fine-tuned alignment.
9. Click the Grid icon again to turn it off.

Once the alignment is done, tighten the C-mount securely into place. As you tighten the screws, the C-mount may move slightly. Make sure that you anticipate that movement as you align the camera.

See Calibration—Troubleshooting

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