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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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Impact of Excess Water Following Eosin

Water is often used as a differentiator for eosin, as it can gently remove excess eosin from the slide. Dehydrating alcohols and the first 100% alcohol can contain enough water to cause lightening of eosin and further differentiation of eosin. Too many "water or diluted alcohol" steps will result in lighter shades of eosin and understaining of cytoplasmic structures.