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Re: beads for Multiview Registration

Posted by Douglas Richardson on Jun 16, 2015; 1:40am
URL: http://imagej.273.s1.nabble.com/beads-for-Multiview-Registration-tp5013158p5013176.html

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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