Method Article

H&E Staining of Paraffin-Embedded Tissue Sections: A Differential Staining Technique to Visualize Liver Tissue Sections Using Combination of Hematoxylin and Eosin Dyes

July 8th, 2025

In This Article

Abstract

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Source: Ellis, J. L et al., Histological Analyses of Acute Alcoholic Liver Injury in Zebrafish. J. Vis. Exp. (2017).

In this video, we describe a technique of hematoxylin and eosin staining of paraffin-embedded tissue sections to study their histology. The procedure includes deparaffinization of the tissue followed by staining the cells in the tissue. Upon staining, the nucleus appears distinctly blue, and the proteins in the cytoplasm display pink coloration.

Protocol

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1. Preparation of staining solutions

  1. Filter Harris hematoxylin stock solution to remove any solids that have precipitated.
  2. Fold a coffee filter in half so that the filter creates a cone. Open the side panel so that the liquid runs through the filter, allowing any debris to be caught.
  3. Place the filter inside a funnel and the funnel into a glass media bottle. Gradually pour the stock hematoxylin through the filter.
    CAUTION: Hematoxylin is hazardous in case of eye contact, ingestion, and inhalation. Always wear a lab coat and gloves and handle the stock solution in a chemical hood.
  4. Prepare....

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Disclosures

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No conflicts of interest declared.

Materials

List of materials used in this article
NameCompanyCatalog NumberComments
95% ethanol (EtOH) Decon Labs, Inc.2801Flammable
Charged slides Fisher Scientific 12-550-16
Dissecting microscope Leica BiosystemsLeica Mz 95
Eosin-Phloxine stain set Newcomer Supply1082A
Ethyl alcohol Sigma-AldrichE7023Flammable
Harris hematoxylin Poly Scientific R&D Corp. s212 Irritant
Hydrochloric acid (HCl) Fisher ScientificA144 Irritant
Isopropanol Newcomer Supply12094EFlammable
XylenesFisher Scientific X3S-4Irritant

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Tags

H E StainingParaffin Embedded TissueHematoxylin Eosin StainingTissue DeparaffinizationTissue RehydrationNuclear StainingCytoplasmic StainingEthanol GradientXylene ClearingMounting Medium Application

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