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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 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.

One Stain Component Overshadowing Another

It is important that the proper balance of the dyes is achieved. Overstaining with hematoxylin can give the illusion of under stained eosin, just as overstaining with eosin can cause the hematoxylin to appear lighter than it actually is. So, when optimizing the stain, make sure to only edit the time of one of the components. This technique will help eliminate the need to spend additional time adjusting the stain.