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The page below is a sample from the LabCE course Theoretical and Practical Aspects of Routine H&E Staining. Access the complete course and earn ASCLS P.A.C.E.-approved continuing education credits by subscribing online.

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Differentiating

As mentioned earlier, differentiation is used with regressive staining techniques and is used to remove the excess dye that has been taken up into the tissue until a crisp delineation of the desired parts is achieved.

Basic dyes are differentiated using a weakly acidic solution prepared with alcohol.

Acid dyes are differentiated using a weakly basic solution prepared with alcohol.

Aqueous based differentiating solutions are not recommended as the dyes will diffuse more rapidly and over-differentiating becomes a problem.