[Histonet] embedding problem small zebrafish

[Andrea Hooper]

Have you checked out the histological methods section of "The Zebrafish Book" by Monte Westerfield, on zfin.org available online here:
http://zfin.org/zf_info/zfbook/cont.html#cont8
I don't think it has paraffin embedding protocols but perhaps there are some other hints will help? In any case, it's an amazing resource.

I am also wondering if it might help to embed in agarose before paraffin embedding - especially those 44 hour old critters who are tiny tiny tiny. Unfortunately most of my experience in that regard lies with frozen sectioning so I don't have too much to add.


----- Original Message -----
From: Barbara Verstraeten <barbara.verstraeten <@t> ugent.be>
Date: Tuesday, March 25, 2008 3:50 am
Subject: [Histonet] embedding problem small zebrafish

> Hello,
> 
> I work on zebrafish from 44 hours to 6 days old (so they are very  
> small; max. 0.5 cm).
> I would like to perform immostainings on them (such as E-cadherin  
> staining). I've embedded my species in technovit 9100 but the 
> cutting  
> of such small animals is very difficult.
> I did some stainings already and the antibody works but the problem 
> is  
> that my tissue is of very bad quality!! When I have 50 sections 
> only 1  
> is usefull.
> This is not very good!
> 
> Can someone give me any suggestions on how to embed them so that 
> they  
> are more easy to section and that I can still stain them with  
> different antibodies?
> 
> Thank you all in advance!
> 
> Kind regards,
> 
> Barbara Verstraeten
> VMDB
> University of Ghent
> Belgium
> 
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