i'm currently trying some H&E staining (post-TUNEL) on paraffin embedded
tissues and keep having the eosin wash out during the dehydration steps
(stepwise: 70% alcohol, 95% alcohol, 100% alcohol)... any ideas on why
this would happen? am i missing a step in between that allows the stain
to 'stick'? thanks
It is 'stuck' but you are unsticking it. Either because you are using
eosin that is soluble in alcohol or because the water in the 70% is
removing the water soluble eosin. You could try adding calcium ions as
an accentuator, that might make the glue stronger, but of course you
must use aqueous eosin or the calcium ions come out of solution.
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