Achieving More Even Lighting

After running all calibrations I always have uneven coloration across my images. This is present even if I move lighting module or redo the flat field correction. The objective is a Plan 60x.

I always seem to have a slightly greener right side and reder left side. The middle has the most saturation. I would love to hear any feedback to get more even lighting.


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Those images I think look really good! Just looking at them on my small screen I can’t really see the colouration that you mention. I’ll try looking in more detail when I can can edit them and lay the left side against the right side to highlight the difference.

I have cropped a part from the right hand side of the first two images and placed it over the left hand side:

There is a very slight change in background colour, I am not sure that it would be possible to do much better than that. The colour of the cells themselves has a bit more difference. That could be more about focus and chromatic aberration. If the slide is not exactly parallel to the camera sensor there will be some focal tilt, and that can also manifest in a slight colour shift.

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Thanks for looking over it. I will try to get a hold of a good objective. It probably is not accurately corrected like a true plan objective.