Dear all,
We are currently facing an issue with our confocal Zeiss LSM 800. Whatever the objective, while a picture is taken, there will always be a dark corner at the bottom right of the image, mostly with the blue and red channels. The attached pictures show both of these channels (DAPI in blue and SOX2 in red), for images taken with the "tiles" tool (x20). You can clealy see that each tile is visibly delimited, which should not be the case, with a dark corner at the bottom right of each tile while the intensity of the rest of the tile is ok (do not mind the oversaturation, it is just to highligh this phenomenon more). Same issue while taking images with the x40 without the tile mode. Slide is properly laying flat on the objective, both are perfectly clean, as well as the camera. Changing intensity parameters or objectives did not resolve this issue. Furthermore each tile has the same profil, indicating that it is not an issue with the slide placement, but rather with the laser position or something like that. Any idea of where this problem is coming from and how to resolve it?
Thank you for your help!
If you sure that "Slide is properly laying flat on the objective, both are perfectly clean, as well as the camera", It may be caused by uneven incident light beam. You can consult an engineer to see if the mercury lamp needs to be adjusted so that the light spot is in the center and evenly distributed.
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