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    The Periodic Acid-Schiff (PAS) Technique

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    vikas
    The periodic acid-Schiff (PAS) technique is the most versatile and widely used method for demonstrating carbohydrates or glycoconjugates in tissues. It involves oxidizing carbohydrates with periodic acid to form aldehydes, which are then demonstrated by reaction with Schiff's reagent to produce a magenta color. The PAS stain labels a wide range of carbohydrates and glycoproteins in tissues, including glycogen, mucins, basement membranes, and fungal capsules. It is a valuable diagnostic tool for evaluating conditions like glycogen storage disease, cancers, infections, and other pathological states.

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    The Periodic Acid-Schiff (PAS) Technique

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    vikas
    320 views18 pages
    The periodic acid-Schiff (PAS) technique is the most versatile and widely used method for demonstrating carbohydrates or glycoconjugates in tissues. It involves oxidizing carbohydrates with periodic acid to form aldehydes, which are then demonstrated by reaction with Schiff's reagent to produce a magenta color. The PAS stain labels a wide range of carbohydrates and glycoproteins in tissues, including glycogen, mucins, basement membranes, and fungal capsules. It is a valuable diagnostic tool for evaluating conditions like glycogen storage disease, cancers, infections, and other pathological states.

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    bmlt pas stain

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    pas-stain

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    The periodic acid-Schiff (PAS) technique is the most versatile and widely used method for demonstrating carbohydrates or glycoconjugates in tissues. It involves oxidizing carbohydrates with periodic acid to form aldehydes, which are then demonstrated by reaction with Schiff's reagent to produce a magenta color. The PAS stain labels a wide range of carbohydrates and glycoproteins in tissues, including glycogen, mucins, basement membranes, and fungal capsules. It is a valuable diagnostic tool for evaluating conditions like glycogen storage disease, cancers, infections, and other pathological states.

    Copyright:

    © All Rights Reserved
    Download as PPTX, PDF, TXT or read online from Scribd
    Download as pptx, pdf, or txt
    320 views18 pages

    The Periodic Acid-Schiff (PAS) Technique

    Uploaded by

    vikas
    The periodic acid-Schiff (PAS) technique is the most versatile and widely used method for demonstrating carbohydrates or glycoconjugates in tissues. It involves oxidizing carbohydrates with periodic acid to form aldehydes, which are then demonstrated by reaction with Schiff's reagent to produce a magenta color. The PAS stain labels a wide range of carbohydrates and glycoproteins in tissues, including glycogen, mucins, basement membranes, and fungal capsules. It is a valuable diagnostic tool for evaluating conditions like glycogen storage disease, cancers, infections, and other pathological states.

    Copyright:

    © All Rights Reserved
    Download as PPTX, PDF, TXT or read online from Scribd
    Download as pptx, pdf, or txt
    of 18

    The periodic acid-Schiff (PAS)

    technique

    The most versatile and


    widely used technique for the demonstration of
    carbohydrates
    or glycoconjugates. The first histochemical
    use of this technique was by McManus (1946)
    for the demonstration of mucin.
    or
    Periodic acid–Schiff (PAS) is a staining method
    used to detect polysaccharides such as
    glycogen, and mucosubstances such as
    glycoproteins, glycolipids and mucins in
    tissues.
    PAS reactive - cell &tissue
     Glycogen
     Starch
     Mucin- sialomucin n neutral mucin
     Alpha- antitrypsin
     Reticulin
     Fungi capsules
     Pancreatic zymogen granules
     Thyroid colloid
     Corpora amylacea & Russell bodies
    PRINCIPLE:
    • The PAS stain is a histochemical reaction in
    that the periodic acid oxidizes the carbon to
    carbon bond forming aldehydes which react to
    the fuchsin-sulfurous acid which form the
    magenta color. ... Use skin, aorta or normal
    liver for positive PAS staining.
    Why does PAS stain basement membrane?

    • The periodic acid-
    Schiff (PAS) staining procedure is most
    commonly used in the histology laboratory to
    detect glycogen deposits in the liver when
    glycogen storage
    disease is suspected. ... Basement
    membranes, which are present in various
    tissues in the body, may also be visualized
    through the PAS staining procedure.
    What Colour does PAS stain?

    • magenta
    • PAS stains carbohydrates and carbohydrate
    rich macromolecules a deep red colour
    (magenta). Mucus in cells and tissues,
    Basement membranes, and Brush borders of
    kidney tubules and small and large intestines
    Reticular fibres (i.e. collagen) in connective
    tissue and Cartilage.
    • CONTROL: For staining fungus; use a known
    positive such as those used for the GMS. Use
    skin, aorta or normal liver for positive PAS
    staining.
    • FIXATIVE: Any well fixed tissue.
    • TECHNIQUE: Cut paraffin sections 4-5 m( 3m for
    kidney biopsies).
    • EQUIPMENT: Rinse glassware in DI water. Coplin
    jar, microwave oven.
    How is Schiff's reagent prepared?
    • Schiff reagent may be made by simply adding
    1 g. fuchsin,.
    • 1.9 g sodium metabisulfite to 100 ml.
    • 0.15 N hydrochloric acid,
    • shaking at intervals or mechanically for 2
    hours,
    • decolorizing with charcoal and filtering.
    . • Caution: Avoid contact and inhalation
    SAFETY:
    • Wear gloves, goggles and lab coat.
    • Avoid contact and inhalation.
    • Hydrochloric acid: strong irritant to skin, eyes
    and respiratory system.
    • Target organ effects via inhalation on skin,
    respiratory, reproductive and fetal systems.
    Corrosive.
    PROCEDURE:
    1. Deparaffinize and hydrate to distilled
    water.
    2. Place slides into 0.5% Periodic acid for 5
    minutes.
    3. Rinse in distilled water.
    4. *Schiff's Reagent, microwave HIGH
    power, for 45 - 60 seconds, until deep
    magenta.
    5. Wash in running tap water for 5 minutes
    6. Counterstain in hematoxylin for 3 minutes.
    7. Wash in tap water, blue hematoxylin, rinse
    in distilled water.
    8. Dehydrate in alcohol, clear, and coverslip.
    *
    Conventional method: Schiff's Reagent, room
    temperature for 30 minutes.
    Result- PAS staining
    A positive test is indicated by a change from pale
    yellow (colourless) to deep red-violet upon
    addition of the aldehyde to the rea- gent.

    Glycogen,neutral/sialomucin,glycoprotein-
    magenta

    Nucleus- blue
    LM- liver;glycogen(PAS)160x
    PM- lung adenocarcinoma(PAS);mucin
    PM- renal corpuscle;PAS

    PM- renal corpuscle;PAS


    Lung- aspergillus;PAS
    PAS stain for glycogen in an alcoholic formalin fixed liver

    Treatment with diastase Without diastase


    Diagnostic utility- PAS
     Glycogen storage disease
     adenocarcinoma
     Paget’s ds- breast
     Alveolar soft tissue sarcoma
     Alpha1 anti-trypsin deficiency
     Mycosis fungoides
     Fungal infection
     Carcinoma of bladder,kidney,liver,ovary,
     Ewings Sarcoma,seminoma,juvenile rhabdomyosarcoma
    • usefulness of PAS staining in identification of
    carbohydrates in normal and malignant lesions of
    mammarygland and endocervical gland

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