Hi,
        I am a novice in the realm of confocal microscopy. I apologize if
some of my questions sound trivial. I am working with mammalian cells and my
first attempt was awesome: staining my cells with a fluorescent phallotoxin
for actin and propidium iodide for DNA (surely well known already to all of
you). I got beautiful well resolved images.
        However, I am now trying other things with disappointing results.
Viability assays as well as staining with labeled antibodies are giving
blurred images, both in fixed and non-fixed preparations. The optical system
is checked, since I saved some of the preps mentioned above and continue to
give good resolution. Is this common? I don't think so, and I have run out
of variables to control and change.
Probably you would like more details about how I worked with this in order
to give suggestions, but anyhow, any ideas are welcome.
        Thanks!
 
                                                           Susi