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Hi Pedro:
If you are using an inverted scope then the areas close to the bottom are less affected by spherical aberration and scattering. That's why they should be brighter.
Mike
________________________________
From: Confocal Microscopy List on behalf of Pedro Camello
Sent: Tue 5/6/2008 6:44 PM
To: [log in to unmask]
Subject: Strange artifact in z series
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Hi all,
while performing z sections of mitochondria-stained living cells, Iīm
finding that the bottom part of the cell displays a brighter mitochondria
fluoresecence, showing more contrast respect to cytosol than mitochondria in
the top part of the cell. My system scans z from bottom to top. If it were
simple bleaching due to laser light passing through the cell, I should
observe that the bottom mitochondria are also less stained in a second
series, but thatīs not the case (the difference between top and bottom still
there)
Any idea to explain it?
Thanks
Pedro
