*****
To join or leave the confocal microscopy listserv or to change your email address, go to:
https://lists.umn.edu/cgi-bin/wa?SUBED1=confocalmicroscopy&A=1
Post images on http://www.imgur.com and include the link in your posting.
*****

Hi Claire

We have the same problem on our LSM980 using Airyscan. Removing the DIC slider out of the light path helped a lot but the PSF is still asymmetric and we don't obtain the advertised best resolution. I did a lot of testing, also together with the Zeiss product specialists, but at the end I was told that all parts are within specifications and this is as good as it gets.

When using Joined Deconvolution in SR-mode the problem is corrected/resolved.

Best wishes
Kees

-----Original Message-----
From: Confocal Microscopy List <[log in to unmask]> On Behalf Of Doug Richardson
Sent: 06 February 2024 01:41
To: [log in to unmask]
Subject: Re: Asymmetric PSF on LSM980 Confocal

***CAUTION:*** This email was sent from an EXTERNAL source. Think before clicking links or opening attachments.

*****
To join or leave the confocal microscopy listserv or to change your email address, go to:
https://lists.umn.edu/cgi-bin/wa?SUBED1=confocalmicroscopy&A=1
Post images on http://www.imgur.com/ and include the link in your posting.
*****

Do they have a DIC slider in the light path?

Get Outlook for Android<https://aka.ms/AAb9ysg>
________________________________
From: Confocal Microscopy List <[log in to unmask]> on behalf of Claire Brown, Dr. <[log in to unmask]>
Sent: Monday, February 5, 2024 8:38:21 PM
To: [log in to unmask] <[log in to unmask]>
Subject: Asymmetric PSF on LSM980 Confocal

*****
To join or leave the confocal microscopy listserv or to change your email address, go to:
https://lists.umn.edu/cgi-bin/wa?SUBED1=confocalmicroscopy&A=1
Post images on http://www.imgur.com/ and include the link in your posting.
*****

Hello All,

I'm posting this for a colleague who is not on the list and is trying to do FCS on a Zeiss LSM980.

"When constructing the PSF to measure its dimensions in the Zeiss LSM 980 using 100 or 20 nm diameter fluorescent beads, there is large astigmatism in the y dimension compared to the x dimension where y is up to 30% larger than x. The same problem is witnessed for the Airyscan PSF on the same microscope. Has anyone else witnessed this? Is that normal? Can that be fixed without impacting the sensitivity or the resolution limits? And how does that impact FCS measurements where the 3D fitting diffusion model assumes x, y symmetry. "


All the best,

Claire