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What are the beads mountd in?  If it's not water the
problem is probably spherical aberration.  Even if
it is water you'll need to adjust the correction collar
to match your coverslip (setting it to the nominal
value is most probably not good enough).  

By eye, not in confocal, adjust the ring till you 
see identical out of focus images above and below
focus.  Then try the XZ series again.

                                                          Guy



----- Original Message ----- 
From: "Sarah Kefayati" <[log in to unmask]>
To: <[log in to unmask]>
Sent: Saturday, July 07, 2007 1:36 AM
Subject: Curvature in YZ plane


> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
> 
> Hello every one,
> 
> It's about two weeks that I've started imaging Tetrascpeck
> beads(.2micron),I see something very strange when I do the sectioning:
> 
> looking at YZ plane, I see the curvy shape ,it's like there is a
> bending point.I thought maybe it's because of astigmatism of our diode
> laser so I switched the laser with Ti-sapphire ,I still can see the
> curvature in yz plane.my second guess is stage displacement in y
> direction during sectioning in z direction,or maybe it's an aberration
> with our objective lens(we're using Olympus 60x/1.2 water immersion
> objective)...
> 
> I appreciate if you have any information regarding this problem and
> share with me.
> 
> regards
> sarah
>