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    Pewarnaan Histologi

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    Stnrhaliza
    This document discusses histological staining techniques used to visualize tissue structures under a microscope. It describes the basic steps of tissue processing which include fixation, dehydration, clearing, infiltration, and embedding. It then explains how different stains can be used to distinguish between tissue components by targeting basophilic, acidophilic, or carbohydrate structures. Common stains discussed include hematoxylin and eosin, PAS, Masson's trichrome, Sudan black, silver stains, and Verhoeff van Gieson. The document also mentions methods to visualize specific molecules using fluorescent, radioactive, enzyme, or metal labels as well as techniques like immunohistochemistry.

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    Pewarnaan Histologi

    Uploaded by

    Stnrhaliza
    79 views16 pages
    This document discusses histological staining techniques used to visualize tissue structures under a microscope. It describes the basic steps of tissue processing which include fixation, dehydration, clearing, infiltration, and embedding. It then explains how different stains can be used to distinguish between tissue components by targeting basophilic, acidophilic, or carbohydrate structures. Common stains discussed include hematoxylin and eosin, PAS, Masson's trichrome, Sudan black, silver stains, and Verhoeff van Gieson. The document also mentions methods to visualize specific molecules using fluorescent, radioactive, enzyme, or metal labels as well as techniques like immunohistochemistry.

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    Pewarnaan Histologi (1)

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    This document discusses histological staining techniques used to visualize tissue structures under a microscope. It describes the basic steps of tissue processing which include fixation, dehydration, clearing, infiltration, and embedding. It then explains how different stains can be used to distinguish between tissue components by targeting basophilic, acidophilic, or carbohydrate structures. Common stains discussed include hematoxylin and eosin, PAS, Masson's trichrome, Sudan black, silver stains, and Verhoeff van Gieson. The document also mentions methods to visualize specific molecules using fluorescent, radioactive, enzyme, or metal labels as well as techniques like immunohistochemistry.

    Copyright:

    © All Rights Reserved
    Download as PPTX, PDF, TXT or read online from Scribd
    Download as pptx, pdf, or txt
    79 views16 pages

    Pewarnaan Histologi

    Uploaded by

    Stnrhaliza
    This document discusses histological staining techniques used to visualize tissue structures under a microscope. It describes the basic steps of tissue processing which include fixation, dehydration, clearing, infiltration, and embedding. It then explains how different stains can be used to distinguish between tissue components by targeting basophilic, acidophilic, or carbohydrate structures. Common stains discussed include hematoxylin and eosin, PAS, Masson's trichrome, Sudan black, silver stains, and Verhoeff van Gieson. The document also mentions methods to visualize specific molecules using fluorescent, radioactive, enzyme, or metal labels as well as techniques like immunohistochemistry.

    Copyright:

    © All Rights Reserved
    Download as PPTX, PDF, TXT or read online from Scribd
    Download as pptx, pdf, or txt
    of 16

    Histological Stains

    Rizka Vidya Lestari


    Department of Histology, Medical Faculty
    Universitas Mataram
    • The most common procedure used in histologic research is the
    preparation of tissue slices or “sections”
    Steps

    • Fixation: To preserve cell and tissue structure (ex: formalin,


    formaldehyde, glutaraldehyde)
    • Dehydration: To removes all water with alcohol solutions
    • Clearing: Alcohol removal with xylol
    • Infiltration: The tissue is placed in melted paraffin
    • Embedding: The paraffin-infiltrated tissue
    • Trimming: Sectioning (slicing) on a microtome
    Staining

    • Most cell and extracellular material are completely colorless


    • Methods of staining can be use to distinguish tissue component from
    one another
    • Dyes stain material often behaving like acidic or basic compounds
    Basic Dyes

    Staining basophilic tissue components (DNA, RNA, and


    glycosaminoglycans) ex:
    • Toluidine blue
    • Alcian blue
    • Methylene blue
    • Hematoxylin
    Acid Dyes

    Stain the acidophilic components


    (mitochondria, secretory granules,
    and collagen) ex:
    • Eosin
    • Orange G
    • Acid fuchsin
    Hematoxylin and Eosin (H&E) is used most commonly. Hematoxylin producing a
    dark blue or purple color, In contrast, eosin producing a pink color

    The PAS-stained tissue was counterstained with hematoxylin to show the cell nuclei
    intensely
    Another Dyes

    • PAS (Periodic Acid–Schiff ): To determine polysaccharides and other carbohydrate


    (purple/magenta)
    • Masson’s trichrome: To distinguish various extracellular tissue components
    • Sudan Black: To determine intracellular accumulations of cholesterol,
    phospholipids, or glycolipids
    • Silver stain/ metal impregnation: Using solutions of silver salts to visual certain
    ECM fibers and specific cellular elements in nervous tissue
    • Verhoeff van Gieson: A special stain for elastic fibers/ connective tissue
    • Giemsa stain: used in cytogenetics and histopathological diagnosis
    • Trypan blue: Direct identification and enumeration of live (unstained) and dead
    (blue) cells
    PAS Masson’s trichrome
    Sudan Black Silver stain
    Verhoeff van Gieson Giemsa stain Trypan blue
    Visualizing Specific Molecules

    • A specific macromolecule may also be identified by using tagged


    compounds that bind specifically with the molecule of interest
    • The most commonly used labels are:
    1. Fluorescent compounds
    2. Radioactive atoms
    3. Enzymes
    4. Metal (usually gold)
    • These methods can be used to detect and localize specific sugars,
    proteins, and nucleic acids
    • Molecules that interact specifically with other molecules:
    1. Phalloidin: Interacts strongly with the actin protein of
    microfilaments
    2. Protein A: Can be used to localize naturally antibodies bound to cell
    structures
    3. Lectins: Bind to carbohydrates with high affinity and specificity
    Immunohistochemistry

    • A highly specific interaction between macromolecules is that between


    an antigen and its antibody
    • To identify and localize many specific proteins
    Enzymes that can be detected
    histochemically:
    1. Phosphatases: Remove phosphate
    groups from macromolecules
    2. Dehydrogenases: Transfer hydrogen
    ions from one substrate to another,
    such as many enzymes of the citric
    acid (Krebs) cycle
    3. Peroxidase: Promotes the oxidation
    of substrates with the transfer of
    hydrogen ions to hydrogen peroxide

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