The H&E Stain: Far From Routine Part 2

This blog is a followup to the previous article “The H&E Stain: Far From Routine”. In that article, the basics of the routine hematoxylin and eosin (H&E) stain were discussed. Now we shall discuss how to trouble shoot the routine H&E and how to ensure a high quality stain, once you have worked with your pathologist to determine the best stain result for your laboratory.

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The H&E Stain: Far From Routine Part 1

What exactly is a routine “H&E”? And what makes it routine? The first question is easy. “H” stands for ”hematoxylin” and “E” stands for “eosin”. Both are dyes used to stain tissue sections in histology. However, the procedure for correctly applying this combination of stains to tissue sections is far from routine.

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Silver Stains

In the histology world, the mere mention of a “silver stain” may be the cause of panic and uncertainty with regard to the performance of the stain, and the quality of the final resulting microscope slide. All other special stains, with few exceptions, are relatively easy and straightforward to perform; not so with silver stains.

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Stains for Microorganisms

The staining of microorganisms in histology can be challenging. Filamentous fungi and associated conidia are more easily demonstrated as they are visible under light microscopy when stained with periodic acid Schiff’s (PAS) as in Figure 1. The diameter of fungi filaments is 5-10 microns, which is approximately the same as the diameter of a red blood cell, while their length may be hundreds of microns (Figure 2). Microorganisms are extremely small and are at the limit of resolution of the light microscope. Viruses are even smaller (Figure 3).

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Staining Fungi

Fungi include molds, yeasts and higher fungi. All fungi are eukaryotic and have sterols but not peptidoglycan in their cell membrane. Their cell walls are composed of cellulose; the same building blocks that plants use.

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Special Stains for Mucins

When staining sections for the presence of carbohydrates, the two main classes under investigation are glycogen and mucins.  Mucins include substances referred to as mucopolysaccharides, mucosubstances, glycoproteins and glycoconjugates.

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