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
Purpose: To troubleshoot a hematoxylin and eosin (H&E) stain with the cytoplasm that is stained too dark.
Specialty: Clinical Laboratory
Objective: laboratory tests
ICD-10: ,