Strange artifact in z series

Posted by Pedro Camello on
URL: http://confocal-microscopy-list.275.s1.nabble.com/Strange-artifact-in-z-series-tp592967.html

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Hi all,

while performing z sections of mitochondria-stained living cells, I´m
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 that´s not the case (the difference between top and bottom still
there)
Any idea to explain it?

Thanks

Pedro