Posted by
Michal Gdula-2 on
URL: http://confocal-microscopy-list.275.s1.nabble.com/Correction-of-Z-axis-distortion-request-for-opinion-tp2580156p2593214.html
Thank you very much for your response!
I scanned slides with 4um beads and the stretching seems to be at least 20%
of the bead. It is hard to say precisely because of blurring.
I checked beads mounted with vectasshield, water and immersion oil. The
biggest extension was in the water, and there was almost no extension in oil
(the spheres looked almost round in xz and yz). I know that it is not anything
new, but I think that it confirms that refraction index is in huge part
responsible for this aberration if not alone.
Best regards,
Michal Gdula
Subject: Re: Correction of Z-axis distortion- request for opinion
From: Guy Cox <
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Reply-To: Confocal Microscopy List
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Date: Sat, 4 Apr 2009 03:10:11 +1100
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Well, the stretching should be 1.518 / 1.457 if simple RI difference is the
cause - ie about 4%. If you are seeing more than that you'd better look
elsewhere - maybe a dodgy Z drive? I suppose changes in SA with depth
would add to the discrepancy, but that depends a lot on how great a depth
you are measuring through. Also, though it's true oil and coverslip should
have the same RI, oil changes with temperature whereas glass doesn't (much)
so if you can use #1.5 and still have enough working distance it might be
better.
Guy