- The alignment might not be perfect, and you might need to adjust the X and Y positions manually with the sliders.
- The alignment might not be perfect, and you might need to adjust the X and Y positions manually with the sliders.
- If one of the channels seems more defocused then the other, adjust the Z direction with the slider.
- If one of the channels seems more defocused then the other, adjust the Z direction with the slider.
- Only change the Rotation if the L shaped object in the middle can not be aligned with changing X and Y directions.
- Only change the Rotation if the L shaped object in the middle can not be aligned with changing X and Y directions.
- At the end, click "Store Calibration" button
- At the end, click "Store Calibration" button
- Keep in mind that the camera alignment will never be perfectly accurate due to chromatic aberrations, and for accurate colocalization studies, you need to perform "Channel Alignment", as described in the following section.