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January 2011

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From:
Ryan Geil <[log in to unmask]>
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Confocal Microscopy List <[log in to unmask]>
Date:
Mon, 24 Jan 2011 16:46:58 -0500
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It is possible to reverse paraffin embedded material back down through your infiltration series and then re-embed in LR White but it is very time consuming and can be hard on the tissue as extra handling and processing are required.  It is much more advisable to start from fresh material and go through a proper LR White infiltration.  That being said, I would recommend both paraformaldehyde and glutaraldehyde for fixation for a good mix of fast penetration and good cross-linking.  I would also use much smaller tissue sections for LR White processing than I would for paraffin embedding.  In my experience, vacuum infiltration is absolutely necessary when working with any leaf tissue.  I did a lot of embedding work in a past life and I very likely have a protocol kicking around somewhere.  If you are interested in having a look, let me know and I will dig it up.
Cheers,
Ryan Geil

On 2011-01-24, at 4:16 PM, annaoru wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
> 
> Hi!
> No I havenīt tried yet but I think I might try just to see the results.
> I havenīt succeeded to find any information regarding this and the
> possibilities around using the same embedding and fixation technique as I
> did prior to Light microscopy but I hope that it will work or that I at
> least can use the same fixation so that the embedded tissue can be melt and
> re-embedded in resin instead. What do you think about that?
> 
> Thank you for helping!
> 
> Regards!
> /Anna
> -- 
> View this message in context: http://confocal-microscopy-list.588098.n2.nabble.com/Embedding-Arabidopsis-Thaliana-for-confocal-tp5954747p5956682.html
> Sent from the Confocal Microscopy List mailing list archive at Nabble.com.

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