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The page below is a sample from the LabCE course Troubleshooting Guidance for Hematoxylin and Eosin (H&E) Stain. Access the complete course and earn ASCLS P.A.C.E.-approved continuing education credits by subscribing online.

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Well-Rounded, Standard H&E - Progressive Protocol

As with the other protocols, I prefer to start off with a basic protocol that is fairly easy to adapt to different pathologists' preferences.
Oven (if on stainer)15 mins
Xylene2 mins
Xylene2 mins
Xylene 2 mins
100% ethanol2 mins
100% ethanol2 mins
95% ethanol1 min
Water wash 1 min
Hematoxylin3 mins
Water wash2 mins
Bluing1 min
Water wash1 min
95% ethanol1 min
Eosin Y45 secs
95% ethanol30 secs
100% ethanol1 min
100% ethanol1 min
Xylene2 mins
Xylene2 mins
Remember, there is flexibility if you would like an additional xylene at the beginning and/or at the end prior to coverslipping. I have also found that this protocol works well with just about any commercially-available hematoxylin or eosin. The protocol is also slightly shorter without any differentiation.