Posted by
Stephen Cody on
URL: http://confocal-microscopy-list.275.s1.nabble.com/Strange-artifact-in-z-series-tp592967p592970.html
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http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocalG'day Pedro,
Are you using a water immersion lens?
A water immersion lens with a coverslip correction collar will be your best chance at minimising the effects of spherical aberration etc. These artefacts will probably still affect your imaging at depth, even with a water immersion lens, but the problem will be greatly reduced.
Cheers
Stephen H. Cody
Microscopy Manager
Central Resource for Advanced Microscopy
Ludwig Institute for Cancer Research
PO Box 2008 Royal Melbourne Hospital
Victoria, 3050
Australia
Tel: 61 3 9341 3155 Fax: 61 3 9341 3104
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-----Original Message-----
From: Confocal Microscopy List on behalf of Pedro Camello
Sent: Tue 5/6/2008 6:44 PM
To:
[hidden email]
Subject: Strange artifact in z series
Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocalHi 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
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