Sebastian is talking about the TILL spinning disc, not the Yokogawa spinning disc. The dichroics will be different between these two devices. With Yokogawa the dichroics is placed between the pinhole disc and the microlens disc and needs to meet specific dimensions, one of those includes a thin substrate on which to sputter you coatings. This thin substrate can be prone to bending, although steps can be taken to minimize this. The TILL system has a single disc and the dichroics is in free space so that you can make it as thick as you choose.
If you would like to discuss further details please contact me offline.
Cheers Robin
Robin Battye, M.Sc., Ph. D.
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-------- Original message --------
Subject: Re: calibration of confocal spinning disk setup (yokogawa)
From: Zac Arrac Atelaz <[log in to unmask]>
To: [log in to unmask]
CC: Re: calibration of confocal spinning disk setup (yokogawa)
*****
To join, leave or search the confocal microscopy listserv, go to:
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Stefan: I am almost sure that Sebastian is talking about a 5mm thick crystal to built the dichroic on to, so all the dichroics will have this thickness. Best regards
> Date: Thu, 28 Jun 2012 15:04:44 +0200
> From: [log in to unmask]
> Subject: Re: calibration of confocal spinning disk setup (yokogawa)
> To: [log in to unmask]
>
> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Hi Sebastian,
>
> I am simply calculating the x/y fwhm of each particle. I already used different objectives, still
> giving the same results, thus it's probably not a matter of the objective. Also we use no
> aftermagnification.
>
> Unfortunately, I don't understand what you mean with using 5mm substrates on the dicroics. Maybe you
> can explain this in more detail?
>
> Thanks!
> Stefan
>
> Sebastian Rhode wrote:
> > *****
> > To join, leave or search the confocal microscopy listserv, go to:
> > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> > *****
> >
> > Hi Stefan,
> >
> > from my experience so far, there can be a couple of things go wrong. When I
> > found asymmetric PSF, the most common problem were caused by:
> >
> > - the objective itself - Did you check the PSF of those beads using the same
> > objective on different setups?
> >
> > - Do you use a CMount with built-in optics (Aftermagnification) since use a
> > CCD with rather large pixels?
> >
> > - Dichroic - especially the CSU-X1 use very thin (0.5mm) dichroics, which
> > will not be very flat (for this reason we use 5mm substrates for our
> > dicroics) - so this might induce a curvature in one direction and finally
> > lead to astigmatism
> >
> > By the way, what are the values you calculated?
> >
> > By the way, no commercial interests here.
> >
> > Cheers, Sebi
> >
> > Dr. Sebastian Rhode
> > Project Manager Research & Development
> >
> > TILL Photonics GmbH
> > an FEI Company
> >
> >
> >
> > On Thu, 28 Jun 2012 12:08:02 +0200, Stefan Sokoll <[log in to unmask]>
> > wrote:
> >
> >> *****
> >> To join, leave or search the confocal microscopy listserv, go to:
> >> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> >> *****
> >>
> >> Hello,
> >>
> >> I am doing single particle tracking using an upright confocal spinning disk
> > setup. The setup
> >> basically consists of an Olympus TIRF100x oil objective, an Olympus
> > microscope and on top a Yokogawa
> >> spinning disk CSU-X1 with an Andor EMCCD camera.
> >>
> >> Analyzing the 3D PSF of latex beats I discover that the PSF is not
> > symmetrical in xy direction,
> >> instead the x expansion is roughly 50nm larger than in y direction.
> >>
> >> I wonder what might be the main cause for this effect? Tilted cover slip,
> > adjustment of the devices,
> >> the CSU itself or ...? So far I just put the devices on top of each other
> > but due to the upright
> >> setup I cannot check the symmetry of the laser beam e.g. over long distance
> > at a wall.
> >> Any ideas on how to best start calibrating the setup are very welcome.
> > Thanks in advance!
> >> Best,
> >> Stefan
> >>
> >> --
> >> ---------
> >> Dipl. Ing.-Inf. Stefan Sokoll
> >> Lab. Molecular Physiology
> >> Dept. Neurochemistry & Molecular Biology
> >> Leibniz Institute for Neurobiology (LIN)
> >> Brenneckestrasse 6
> >> 39118 Magdeburg
> >> Germany
> >>
> >> Mail: [log in to unmask]
> >> Tel.: +49 - 391 - 626393171
>
> --
> ---------
> Dipl. Ing.-Inf. Stefan Sokoll
> Lab. Molecular Physiology
> Dept. Neurochemistry & Molecular Biology
> Leibniz Institute for Neurobiology (LIN)
> Brenneckestrasse 6
> 39118 Magdeburg
> Germany
>
> Mail: [log in to unmask]
> Tel.: +49 - 391 - 626393171
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