A Good Baseline Protocol

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A Good Baseline Protocol

A mid-range stain is often preferred. The protocol below is utilized by the author when setting up a new laboratory or when helping to optimize an existing stain that has become less then optimal.
Reagent / Step Time
Xylene 2 minutes
Xylene 2 minutes
Xylene2 minutes
100% ethanol2 minutes
100% ethanol 2 minutes
95% ethanol2 minutes
Water wash2 minutes
Hematoxylin3 minutes
Water wash1 minute
Differentiator (mild acid) 1 minute
Water wash1 minute
Bluing1 minute
Water wash1 minute
95% ethanol1 minute
Eosin1 minute
95% ethanol1 minute
100% ethanol1 minute
100% ethanol1 minute
Xylene1 minute
Xylene2 minutes
Xylene 2 minutes
Coverslip