Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

The problem consist in getting not-so-well-focused images on screen, while
the cell viewed by eye appears super sharp. We tested it on
phalloidin-labelled cells: we see sharp stress fibres and striations in
cardiomyocytes and the same stress fibres appear slightly blurred on the
monitor screen and striations are hard to make up.

Any advice on what to adjust before reaching for the wallet?



Michal,
        You don't mention how you are collecting the images and what is
occurring during the scan process.  There are several things that can cause
this, most of which have to do with dirty contacts on the connection from
the PMT's to the AD converters.  Do you see a wave appearance in the scan if
you Kalman average?  That is there an image shift from one scan to the next.
This will cause blurring.  It appears as a wave from one scan to the next
and will be most apparent at scan speed 1 ( slow)  To remedy this remove the
front panel.  You have to remove the top plate or end plate if on an
inverted.  Slide the cover off and remove the 9 pin cables running into the
converter. These are located on the left side if system is upright   Clean
end with a clean pencil eraser.  I sometimes use electronic cleaner as well.
Also remove the 12-16  pin connector cables in the middle and clean these as
well.  Then be sure all your mirrors are aligned.  I suspect the problem
will go away then.  I usually do this 2-3 times a year depending upon usage
level and it has always worked for me.  You may have to do the same to the
cables running from the scan head to the frame store junction box.

Joe Goodhouse
Confocal / EM Core Laboratory
Department of Molecular Biology
Princeton University
609-258-5432

Visit us at  http://www.molbio.princeton.edu/facility/confocal/index.html