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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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The hematoxylin stain in this section of colon appears to be taking over
the section. Even the mucin in the goblet cells are stained blue.

The nuclear stain in this kidney section is very light.
Simply increasing the time in hematoxylin by 30 seconds to one minute
may be enough to darken the nuclei enough for diagnosis.

Is the Differentiator the Right Intensity?

With regressive and modified progressive staining, a differentiator is used. If the differentiator is made in-house, there is the potential for it to be either too weak or too strong. Both scenarios will impact staining. If the differentiator is stronger than intended, it will remove more hematoxylin and will make the nuclei pale. Time is also important. Too much time in a properly prepared differentiator will also remove more hematoxylin and will ultimately under stain the nuclei.