Hello,
We have a Zeiss Axio Examiner microscope which we use for ex vivo electrophysiology. The microscope has a 10X objective and a 63X objective, both mounted on a rolling slide.
After removing and putting back the 63X objective for some routine cleaning, we realized that the two objectives don't exactly focus on the same point of the specimen. Notably, if we focus on a specific region of the sample with the 10X, there is a high chance that, when switching to the 63X, the image taken will be outside of the slice.
Looks like a problem of diffraction or optical aberration to me, does it make sense? What could be a solution to this?
Did anybody experienced something similar with in vivo or in vitro microscopy?
Thank you very much for your help!
Hi Luca,
From what I understand, the problem is that when you switch between the two objectives, the field-of-view may sometimes change. However, this does not happen all the time.
First I want to rule out some simple errors that may cause this.
1) When you screw the objectives back in, did they go in straight and smoothly? Sometimes one can screw threads in a little bit at an angle.
2) When switching between the objectives, did you make sure that the slider is completely in position of the slots?
3) When switching between the objectives, could it be possible that the stage was moved accidentally?
Let me know if these already helped, or maybe there is indeed a more serious problem with the light path.
Best,
Aaron
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