Re: Fluorescent grid (in z) slide

Posted by zhouzc on
URL: http://confocal-microscopy-list.275.s1.nabble.com/Fluorescent-grid-in-z-slide-tp7591970p7591983.html

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Hi all,

Great ideas - I'm looking into the argolight slides and I think your solution Zdenek will be a nice interim method.

Thanks all!
Charles

-----Original Message-----
From: Confocal Microscopy List <[hidden email]> On Behalf Of Zdenek Svindrych
Sent: Friday, March 12, 2021 8:42 AM
To: [hidden email]
Subject: Re: Fluorescent grid (in z) slide

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Hi Steffen,
one use case where z-calibration is non-trivial is when the GRIN is fixed to the sample and you're focusing with a movable objective lens on the other side of the GRIN (or more exotic arrangements).
Also, GRIN lenses (at least those I worked with) cannot be made telecentric, so the PSF is not constant over the field of view...(pain)

Charles:
For the z-calibration I was thinking about some layers of tape (or thin
coverslip) with some beads in between, but the z-spacing will be rather coarse (50 - 100 um at least) and not very uniform.

Besty, zdenek

On Fri, Mar 12, 2021 at 3:36 AM Steffen Dietzel <[hidden email]> wrote:

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>
> the Argolight slide has a sphere made of 3 rings: one in xy, one in xz
> and one in yz. Also, there are several z-stair patterns on it, with
> different z-step sizes. It sounds like you would prefer a cube with a
> 3D-dot pattern (which might be a good idea for future versions) but
> maybe one of those patterns would do. argolight.com
>
> It is not cheap, but it gives you a lot of options for quality control.
> If you should have money to burn, you might get a custom pattern. No
> commercial interest on my side.
>
> Out of curiosity. should you not be able to see 3D distortion if you
> focus through a 2D pattern? Not that I would know much about grin
> lenses. At the end of the day, every available detector just takes a
> 2D image at max, and assembles 3D in the computer.
>
> Steffen
>
>
> Am 11.03.2021 um 17:39 schrieb zhouzc:
> > *****
> > To join, leave or search the confocal microscopy listserv, go to:
> > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> > Post images on http://www.imgur.com and include the link in your
> posting.
> > *****
> >
> > Hi all,
> >
> > I'm trying to measure how grin lenses distort and magnify the image
> > in the z-axis. I know there are 2D fluorescent grids that are evenly
> > spaced (eg. https://www.psfcheck.com/about-us) for lateral
> > measurements, but I was wondering if there any similar
> > slides/samples but for axial measurements.
> >
> > Thanks!
> > Charles
>
> --
> ------------------------------------------------------------
> Steffen Dietzel, PD Dr. rer. nat
> Ludwig-Maximilians-Universität München Biomedical Center (BMC) Head of
> the Core Facility Bioimaging
>
> Großhaderner Straße 9
> D-82152 Planegg-Martinsried
> Germany
>
> http://www.bioimaging.bmc.med.uni-muenchen.de
>

--
--
Zdenek Svindrych, Ph.D.
Research Scientist - Microscopy Imaging Specialist Department of Biochemistry and Cell Biology Geisel School of Medicine at Dartmouth