eoe sub articles

EoE Sub Articles

1. Preparation of staining solutions
<ol>
	<li>Filter Harris hematoxylin stock solution to remove any solids that have precipitated.</li>
	<li>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.</li>
	<li>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...

h&amp;e staining of paraffin-embedded tissue sections: a differential staining technique to visualize liver tissue sections using combination of hematoxylin and eosin dyes

Source: Ellis, J. L et al., <a href="https://www.jove.com/v/55630">Histological Analyses of Acute Alcoholic Liver Injury in Zebrafish.</a> 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.

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

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 ...

Begin Hematoxylin and Eosin staining by taking a glass slide containing the paraffin-embedded tissue section. Dip the slide in a deparaffinization solution containing xylene - an organic solvent that dissolves the paraffin layer around the tissue, making it accessible to stains in subsequent steps.
Treat the tissue section with absolute alcohol to remove the xylene. Immerse the tissue in decreasing concentrations of alcohol. This step rehydrates the tissue by replacing the alcohol with water for better compatibility with water-soluble dyes.
Stain the tissue with basic hematoxylin, which in its oxidized form binds to mordant - a metal cation. This complex electrostatically binds to the nucleic acids in the genome, staining the nucleus purple.
Submerge the stained section in an acidic differentiation solution to remove excess hematoxylin from the cytoplasm, making the dye-retaining nucleus appear distinct. Treat the tissue with an alkaline bluing agent to neutralize the free acids. This regresses the purple dye color to blue, enhancing the nuclear details.
Immerse the slide in acidic eosin solution to stain the basic protein components in the cytoplasm and the connective tissue fibers in shades of red to pink. Treat the tissue with alcohol to eliminate excess eosin. Use increasing concentrations of ethanol to dehydrate the tissue. Dip the slide in xylene to clear any remaining reagents.
Upon imaging, the cells in the tissue section show clear blue nuclei and pink cytoplasmic regions.

he-staining-paraffin-embedded-tissue-sections-differential-staining

Research

JoVE Encyclopedia of Experiments

Biological Techniques

Staining Techniques

Tissue staining

Sectioning Mammary Gland Whole Mounts for Lesion Identification

Normal mammary gland development may be altered by exposure to environmental toxicants and pharmaceutical products, excessive exposure to hormones, and genetic alterations. Mammary gland whole mounts are an inexpensive method to capture the progression of morphological changes that may arise after exposure. However, in later life, when abnormalities are more prone to develop, sole reliance on this one method may not always provide enough information to make a proper diagnosis of the abnormality. Historically, in chemical test guideline studies, a single mammary gland is removed at necropsy and prepared as a hematoxylin and eosin (H&#38;E)-stained section. The incorporation of contralateral mammary whole-mount collection and analysis decreases the likelihood of a false-negative assessment. Evaluation of the whole mount is limited by the presence of one or two entire mammary glands on a slide, and in some cases, the abnormalities observed in the whole mount are not uniformly represented in the H&#38;E section.&#160; The goal of this study was to develop a protocol for converting coverslipped mammary whole mounts to H&#38;E-stained sections so that lesions that would otherwise have been missed or that are difficult to diagnose can be identified. Here, we detail a method to produce a high-quality, paraffin-embedded H&#38;E section from a mammary gland that was initially prepared as a whole mount. In comparison to a tissue that was intentionally prepared for H&#38;E sectioning, the whole mount requires additional preparation for tissue removal and processing. However, this method is considered inexpensive, as it requires common lab reagents and little additional time. As a result, this method can provide invaluable information on how chemical and environmental exposures alter normal mammary development, as well as display changes that occur because of genetic modifications.

We developed a method to successfully remove, process, section, and stain, for histopathological evaluation, mammary tissue that had originally been fixed on slides as whole mounts. This method may promote the collection and evaluation of mammary gland whole mounts in reproductive and developmental test guideline studies.

Normal mammary gland development may be altered by exposure to environmental toxicants and pharmaceutical products, excessive exposure to hormones, and ...

JoVE Journal

Cancer Research

Elastic Staining on Paraffin-embedded Slides of pT3N0M0 Gastric Cancer Tissue

The elastic lamina, which is usually located in the sub-mesothelial layer, adjacent to the peritoneum mesothelial cells, is amphiphilic on hematoxylin and eosin (H&amp;E) staining but can be visualized through elastic staining. One of the important benefits of elastic staining is that it makes it easy to determine whether tumor cells have invaded beyond the elastic lamina. This helps in examining the extent of peritoneal surface invasion, thereby distinguishing the pT3 and pT4 stages in gastrointestinal cancer. In this study, we present a protocol to identify elastic fibers in the formalin-fixed, paraffin-embedded pT3N0M0 gastric cancer tissue sections. We prepare 5-&#181;m paraffin sections fixed on slides, and then all the slides are deparaffinated and rehydrated during the staining procedure. Subsequently, all the sections are oxidized by potassium permanganate, bleached by oxalic acid, stained by elastic staining, and counterstained by Van Gieson's. Finally, we examine the effect of staining; elastic lamina is stained blue-black and collagen fibers are stained red, while the cancer cells are stained in varying shades of yellow. We also describe in detail the methods used for determining the positional relationship between cancer cells and elastic lamina. This method is simple, low-cost, and widely applicable in the identification of peritoneal surface invasion in gastrointestinal cancer because of its outstanding selectivity for elastic fibers and authentic results.

Here, we present a protocol of elastic staining to identify elastic fibers in pT3N0M0 gastric cancer tissues on formalin-fixed, paraffin-embedded sections. Subsequently, we describe a method to determine whether tumor cells invade beyond the elastic lamina.

The elastic lamina, which is usually located in the sub-mesothelial layer, adjacent to the peritoneum mesothelial cells, is amphiphilic on hematoxylin and ...

qPCR Is a Sensitive and Rapid Method for Detection of Cytomegaloviral DNA in Formalin-fixed, Paraffin-embedded Biopsy Tissue

It is crucial to identify cytomegalovirus (CMV) infection in the gastrointestinal (GI) tract of immunosuppressed patients, given their greater risk for developing severe infection. Many laboratory methods for the detection of CMV infection have been developed, including serology, viral culture, and molecular methods. Often, these methods reflect systemic involvement with CMV and do not specifically identify local tissue involvement. Therefore, detection of CMV infection in the GI tract is frequently done by traditional histology of biopsy tissue. Hematoxylin and eosin (H&#38;E) staining in conjunction with immunohistochemistry (IHC) have remained the mainstays of examining these biopsies. H&#38;E and IHC sometimes result in atypical (equivocal) staining patterns, making interpretation difficult. It was shown that quantitative polymerase chain reaction (qPCR) for CMV can successfully be performed on formalin-fixed, paraffin-embedded (FFPE) biopsy tissue for very high sensitivity and specificity. The goal of this protocol is to demonstrate how to perform qPCR testing for the detection of CMV in FFPE biopsy tissue in a clinical laboratory setting. This method is likely to be of great benefit for patients in cases of equivocal staining for CMV in GI biopsies.

This protocol describes qPCR detection of cytomegalovirus in formalin-fixed, paraffin-embedded biopsy tissue, which is rapid, sensitive, specific, and useful for interpreting equivocal hematoxylin and eosin or immunohistochemical staining patterns.

It is crucial to identify cytomegalovirus (CMV) infection in the gastrointestinal (GI) tract of immunosuppressed patients, given their greater risk for ...

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

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