Hi.
First of all I would like to thank all of you for your advices.
Because, my previous e-mail wasn't complete and to avoid further
suggestions regarding protocols I can not follow, here I write some
more information.
First of all, we can't perfuse our rats. Also, when we harvest rat
spinal cord (cut in half, thus around 3mm diameter) we fix it only for
4-5 hours in 4% paraformaldehyde because we need to perform an IHC
essay so we would like to avoid epitope masking.
We don't think fixation is the problem since frozen sections of the
same fixed spinal cords do not show any problem in the white matter.
In conclusion, we need to obtain good results using paraffin both in
term= s of morphology and IHC staining.
Our paraffin embedding machine program is as follows:
Ethanol 95% 1h
Ethanol 95% 45'
Ethanol 95% 45'
Ethanol 100% 45'
Ethanol 100% 1h
Ethanol 100% 1h
Ethanol 100% 1h
Xylol 45'
Xylol 1h
Xylol 1h
Paraffin 1h
Paraffin 1h
Paraffin 1h
Can you tell me if are there any specific protocols to embedd rat
spinal = cord in paraffin to avoid washing out of myelin?
Thank you.
Regards
--
--
Dott.ssa Elisa Ballarini,PhD student
Dipartimento di Neuroscie= nze e Tecnologie Biomediche
Università degli Studi Milano-Bicocca
Via Cadore 48-20052,Monza,MB
e.ballari...@campus.uni= mib.it
Tel. 02-64488119
Fax. 02-64488253
--- segue il m= essaggio inoltrato ---
_______________________________________________
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
