H and e staining pdf

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h and e staining pdf

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Most cells are colourless and transparent, and therefore histological sections have to be stained in some way to make the cells visible. The techniques used can either be non-specific, staining most of the cells in much the same way, or specific, selectively staining particular chemical groupings or molecules within cells or tissues. Staining usually works by using a dye, that stains some of the cells components a bright colour, together with a counterstain that stains the rest of the cell a different colour. Acidic dyes react with cationic or basic components in cells.

Routine H&E Staining

Staining of the tissue sections is carried out to study the Morphological and relationship of tissues and their constituent cells. Various types of dyes either Natural or Synthetic, are used to prepare the staining solutions. In histopathology laboratory, the Hematoxylin and Eosin stain is referred as the Routine Stain as it can be used to stain any tissue specimen to reveal the underlying tissue structures and conditions. Eosin is most suitable combination used with the Hematoxylin for the routine staining purpose. Eosin Yellow Y is the most commonly used stain in histopathology laboratory.

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Alejandro S. Tissue staining during a frozen section is one of the most important steps in obtaining an accurate pathologic diagnosis. Visualization and discernment of microscopic details can become difficult if staining is done improperly. The two main dye components are hematoxylin and eosin. It is a basic dye that stains acidic cell components such as nucleic acids, glycosaminoglycans, and acid glycoproteins, into a blue-purple hue.


This stain has been unchanged for many years because it works well with a variety of fixatives and displays a broad range of cytoplasmic, nuclear, and extracellular matrix features. So far it is still the most widely used stain in medical diagnosis and is often the gold standard. Hematoxylin is a compound extracted from the heartwood of the logwood tree. Haematoxylin can be considered as a basic dye. It is used to stain acidic structures a purplish blue.

This uses a combination of two dyes, Hematoxylin and Eosin used for demonstration of nucleus and cytoplasmic inclusions in clinical specimens. Alum acts as mordant and hematoxylin containing alum stains the nucleus light blue. This turns red in presence of acid, as differentiation is achieved by treating the tissue with acid solution. Bluing step converts the initial soluble red color within the nucleus to an insoluble blue color. The counterstaining is done by using eosin which imparts pink color to the cytoplasm. Hai i am lab technician , and also bsc microbiology i like your artile from many technician technician gain knowledge.

Hematoxylin and Eosin Stain. (H&E). 5 micron Paraffin Sections. PROCEDURE: 1. Deparaffinize and rehydrate slides to distilled water. 2. Stain in Mayers.

H&E Staining Overview: A Guide to Best Practices

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We continue to use this simple and essential stain today, which has remained unchanged for well over a century. If you have viewed this educational webinar, training or tutorial on Knowledge Pathway and would like to apply for continuing education credits with your certifying organization, please download the form to assist you in adding self-reported educational credits to your transcript. The format is easily reproduced and the reagents resilient enough to allow for large numbers of slides to be stained consistently before reagents need to be changed. In the past, stains and their components were routinely made by the laboratory. Commercially available, premade reagents were uncommon and expensive.

Staining Methods in Frozen Section: Best Lab Practices

Most dyes used to visualize the membranes and organelles of the cell are water soluble.

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H&E Staining Overview: A Guide to Best Practices

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  • The staining procedure for H&E follows a basic protocol: Dewaxing; Dehydration; Hematoxylin; Differentiation; Bluing; Eosin; Dehydration; Clearing. Albracca F. - 13.05.2021 at 16:16