[Histonet] unsolved proplem

[Jennifer MacDonald]

There are a couple of things that it might be. 
1.   Uneven deparaffinization before staining.
2.  Water in your last reagents/paraffin on the processor.



From:   mohamed abd el razik via Histonet 
<histonet at lists.utsouthwestern.edu>
To:     "Histonet at lists.utsouthwestern.edu" 
<Histonet at lists.utsouthwestern.edu>
Date:   01/26/2016 11:33 AM
Subject:        [Histonet] unsolved proplem



Dear allI have submitted a proplem about reprocessing tissue blocks that 
have patches of stained areas and unstained pale yellow areas (H&E 
stain)!!!, unfortently the proplem didn't solved yet and i have tried all 
possible solutions, increasing processing time and ordered new chemicals 
from other soruces and changed the paraplast company but still have the 
proplem !!!! 
i'm using 70% alc 2hr - 80% alc 1hr - 90% alc 45mins- absolut 1 and 2 each 
for 30 mins then Benzen 1 and 2 each for 25 mins then clearing by methyl 
benzoat 2hs at least then paraffin1+2 (Maccormick melting degree 
56-58)each 90 mins for mice kidney, liver and tests
the previous protocol was very satisfactory to us and we have tried to 
stain older sections processed befor the emerging proplem and have a nice 
stain by current H&E stain to ensure that the proplem is out our staining 
step. 
any suggestions please !!
Mohamed


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