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HEMATOXYLIN AND EOSIN (H&E)

Used for elucidation of basic histologic features, prior to the use of special stains or immunohistochemical studies as needed; among other features, calcification and microorganisms such as fungi and bacteria may be detected by H&E and confirmed by additional studies

STAINS FOR CARBOHYDRATES

  • Periodic-acid Schiff (PAS)

    • Stains glycogen red—diastase labile; therefore, diastase pretreatment will remove glycogen

      • Stains mucopolysaccharides red—diastase stable

      • Stains fungi red—diastase stable

      • Stains basement membrane red—diastase stable

    • Colloidal iron

      • Stains mucin blue

    • Alcian blue

      • Stains mucopolysaccharides blue

        • – At pH 2.5: acid (carboxylated or sulfated mucopolysaccharides)

        • – At pH 1.0: acid (sulfated mucopolysaccharides)

        • – With hyaluronidase: only epithelial mucins will stain (connective tissue mucins will be digested and will not stain)

      • With PAS: acid mucopolysaccharides will stain blue and neutral polysaccharides will stain magenta; also, the yeast of Cryptococcus will stain red and the capsule will stain blue with this method

    • Mucicarmine

      • Stains epithelial mucins red (also stains capsule of Cryptococcus red)

STAINS FOR PIGMENTS

  • Fontana-Masson (Fig. 30-1)

    • Stains melanin and argentaffin granules black (nuclei will be red); useful for quantifying melanocytes (e.g., in vitiligo) and in cases of minocycline pigmentary alteration

    • Also stains Cryptococcus and the phaeohyphomycotic organisms

  • Grimelius argyrophil stain

    • Argentaffin and argyrophil substances will stain black

  • Tyrosinase (DOPA-oxidase)

    • Requires fresh tissue

    • Stains melanin-containing cells brownish-black (due to tyrosinase acting on DOPA, the substrate for this reaction)

FIGURE 30-1

Fontana-Masson stains melanin black in this biopsy of minocycline pigmentation (200x) (also see Fig. 30-3).

STAINS FOR MINERALS

  • Von-Kossa

    • Stains calcium salts black; useful for detecting calcification of vessel walls and elastic tissue (calcinosis cutis, pseudoxanthoma elasticum, calciphylaxis, elastosis, and elastofibroma)

  • Alizarin red S

    • Stains calcium red

  • Prussian blue stain (Fig. 30-2)

    • Stains iron blue (the Prussian blue reaction: tissue treated with dilute hydrochloric acid and potassium ferrocyanide) (Note: if it's granular and staining blue, it's likely an iron stain)

  • Gomori methenamine silver (GMS) (Fig. 30-3)

    • Stains urates (e.g., gout) black. (Note: urates are lost if tissue is processed in formalin)

    • Tissue must be processed with alcohol to prevent loss of urates

    • Also see section "Stains for Microorganisms"

FIGURE 30-2

PAS stain showing dermatophytes in stratum corneum (200x).

STAINS FOR CONNECTIVE TISSUE COMPONENTS

  • Trichrome

    • Stains collagen blue or green and muscle red, depending on the type of reagents used

  • Verhoeff-van Gieson

    • Stains elastic fibers black, collagen red, and muscle yellow (also red cells will stain yellow)

  • Acid-Orcein

    • Stains elastic fibers brown. When combined with giemsa, stains amyloid "sky-blue"

  • Gomori aldehyde fuchsin

    • Elastic fibers are stained purple

STAINS FOR AMYLOID

  • Congo red

    • Stains ...

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