[Histonet] Eosin not working for xylene free H&E

[Margaret Horne]

We don't have problems with our eosin after Xylene-free deparaffining. I
use a 95% alcohol step just before the Eosin. Perhaps that might help.
 
We don't air dry before coverslipping as my prof finds the quality not
as good, so we use 100%IsOH before coverslipping; the mounting media you
use will determine if it is ok to cover slip from IsOH.
 
 
We just had a grad student compare Hot IsOH vs mineral oil vs Hot
detergent as xylene free alternatives.  I'll send you our final protocol
if you like.
 
            
Margaret Horne
AVC, UPEI,
 
 
 
 
 "Gerard Spoelstra" <G.Spoelstra <@t> murdoch.edu.au> 27/03/2013 11:06 AM
>>>
Hi everyone,

I have not had any success optimising the eosin counterstain in the
absence of xylene. We routinely use 0.5% eosin Y in 95% ethanol. I've
tried taking sections through to this to absolute ethanol but then air
drying , but it leaves blotches of eosin.  Using eosin 0.5% aqueous with
a few drops of acetic acid doesn't appear to work, the keratin stains
heavily but not elastin. After washing in water I leave for a short time
in the oven then at room temperature for 5 minutes then coverslip with
the dako atomatic cover slipper.   For all the laboratories that have
gone xylene free, are you just putting up with less than optimal eosin
staining?

Gerard Spoelstra  
Murdoch University
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