Hi Everyone,
I am working on standardising a protocol for mouse EB histology. Currently I have tried various methods to make sure that my EBs are placed in one spot in the embedding wax, but I have always lost majority of the EBs when transferring from xylene to wax. I tried searching for various protocols w.r.t Embryo and EB processing but many of the protocols do not mention in detail how they performed this particular step. Since I do not have any of the latest equipments to work with, I would really appreciate some suggestions which involve using basic histological techniques to process EBs.
Thank you!!!
The routine labe we arer working with embedds small tissues in agar befor paraffinization. maybe that's a method worth looking into for you.
It is something similar to this:
http://www.palpath.com/MedicalTestPages/agarrecipe.htm
Thank you Victoria.! This technique looks promising. I will try it out on a batch of EBs.
Hi Bhushan, I agree with Victoria. The best way to save small tissue pieces during embedding is to embed them at first in agar.
Here my protocol: Prepare a 2 % agar solution. After you have solved the agar in hot water cool it down to 60°C. Prepare a small tissiue block and embed this block in your noraml embedding protocol. You will see when you transfer this block from xylene into paraffin and even when you prepare your paraffin block it is very easy to orientate this block. Don't warry the agar does not interfere with staining protocols.
Good luck!
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