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

[Gerard Spoelstra]

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