beads for Multiview Registration

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beads for Multiview Registration

Mario Emmenlauer-3
Dear All,

for SPIM registration with the Multiview Registration plugin, what beads
size/color/concentration, and imaging settings are people using? We're
interested in time series of Zebrafish, Drosophila and others, so all
input is welcome!

Thanks and all the best,

    Mario


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Balanstrasse 43                    mailto: mario.emmenlauer * unibas.ch
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Re: beads for Multiview Registration

Stephan Janosch
Hey Mario,


we use
http://www.emdmillipore.com/US/en/product/Fluorescent-Functionalized-Microspheres---F1-Z-030,MM_NF-39420083?bd=1

with a final concentration of 0.0005 %

And for imaging L1 c.elegans we usually go to full zoom with the LZ 1. I
hope this helps.

Working with 0.0004 % concentration resulted in difficult registration
situation.

Hope this helps and mybe Stephan Preibisch can put his concentrations
for from his early worm imaging.
Stephan

ps. Christopher Schmied might also have valuable infos fro you.

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Stephan Janosch
Software Engineer - TransgeneOme Database
https://transgeneome.mpi-cbg.de

Max Planck Institute of Molecular Cell Biology and Genetics
Pfotenhauerstr. 108
01307 Dresden
Germany

Room: 205
Phone: +49 351 210 2709
Email: [hidden email]
Web: www.mpi-cbg.de
Twitter: https://twitter.com/TransgeneOme

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Re: beads for Multiview Registration

Douglas Richardson
We've tested a number of different sizes.  We find 200nm beads work best
with our 20x/1.0 objective (we order fluorospheres from Life Technology and
use at a dilution of 1:500 000), but we need to go to 1um beads with our
5x/0.16 objective. These have to be used at a higher concentration, usually
1:2000 dilution from stock.

Personally I prefer 100nm beads for use with the 20x (as these are usually
too dim to be seen in the final reconstruction), but the processing
requires an extra step.  Before registration, I adjust the min/max values
to completely saturate the sample (becomes a giant blob) and allow easy
visualization/registration of the beads).  I convert this to 8bit and
perform the registration.  I then use these registration files on the
original data.

Hope this helps!

-Doug

On Mon, Jun 15, 2015 at 6:53 AM, Stephan Janosch <[hidden email]> wrote:

> Hey Mario,
>
>
> we use
>
> http://www.emdmillipore.com/US/en/product/Fluorescent-Functionalized-Microspheres---F1-Z-030,MM_NF-39420083?bd=1
>
> with a final concentration of 0.0005 %
>
> And for imaging L1 c.elegans we usually go to full zoom with the LZ 1. I
> hope this helps.
>
> Working with 0.0004 % concentration resulted in difficult registration
> situation.
>
> Hope this helps and mybe Stephan Preibisch can put his concentrations
> for from his early worm imaging.
> Stephan
>
> ps. Christopher Schmied might also have valuable infos fro you.
>
> --
> Stephan Janosch
> Software Engineer - TransgeneOme Database
> https://transgeneome.mpi-cbg.de
>
> Max Planck Institute of Molecular Cell Biology and Genetics
> Pfotenhauerstr. 108
> 01307 Dresden
> Germany
>
> Room: 205
> Phone: +49 351 210 2709
> Email: [hidden email]
> Web: www.mpi-cbg.de
> Twitter: https://twitter.com/TransgeneOme
>
> --
> ImageJ mailing list: http://imagej.nih.gov/ij/list.html
>

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