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

Abstract

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

1. Preparation of staining solutions Filter Harris hematoxylin stock solution to remove any solids that have precipitated. 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. 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…

Materials

95% ethanol (EtOH)	Decon Labs, Inc.	2801	Flammable
Charged slides	Fisher Scientific	12-550-16
Dissecting microscope	Leica Biosystems	Leica Mz 95
Eosin-Phloxine stain set	Newcomer Supply	1082A
Ethyl alcohol	Sigma-Aldrich	E7023	Flammable
Harris hematoxylin	Poly Scientific R&D Corp.	s212	Irritant
Hydrochloric acid (HCl)	Fisher Scientific	A144	Irritant
Isopropanol	Newcomer Supply	12094E	Flammable
Xylenes	Fisher Scientific	X3S-4	Irritant