The Hematoxylin and Eosin (H&E) stain is a commonly used combination in histopathology. Too much eosin can result in cytoplasmic staining that is too dark.
Problem
Solution
eosin concentration is too high
Use the correct eosin concentration.
eosin stain also contains phloxine
Reduce the concentration of eosin or change to a formulation without phloxine.
time in eosin solution was too long
Reduce the time in the eosin solution.
inadequate rinsing in alcohol after the eosin step (differentiation)
Make sure that the rinsing in alcohol is adequate. Change the alcohol solutions frequently to minimize contamination.
use of an alcohol other than ethanol to rinse the slide after the eosin step
Use ethanol to rinse the slide.
use of an aqueous eosin solution
Reduce time in the eosin solution or change to an alcoholic eosin.
use of an alcoholic eosin AND slide not adequately dehydrated prior to staining
Make sure that the slide is adequately dehydrated prior to the eosin step. Do not use isopropyl alcohol just prior to the eosin step.
use of an alcoholic eosin AND eosin stain is contaminated with water
Change the eosin solution.
where:
• Phloxine is a xanthene dye referred to as Acid Red 92 or Eosin 10B
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