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  • Stain Techniques

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    rafael santamaría
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    Staining techniques are used to diagnose tissue samples under the microscope. The main techniques are hematoxylin and eosin staining, special stains, immunohistochemistry, and in situ hybridization. Hematoxylin and eosin staining uses dyes to differentiate cell structures and can provide a final diagnosis in 70-80% of cases. More advanced techniques like immunohistochemistry use antibodies to identify proteins, while in situ hybridization identifies DNA and RNA, assisting in diagnosis of the remaining cases. Together these staining methods are essential for pathologists to examine tissue at a microscopic level and determine if a sample is benign or malignant.

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    Técnicas de tinción

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    Stain techniques.pptx

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    Staining techniques are used to diagnose tissue samples under the microscope. The main techniques are hematoxylin and eosin staining, special stains, immunohistochemistry, and in situ hybridization. Hematoxylin and eosin staining uses dyes to differentiate cell structures and can provide a final diagnosis in 70-80% of cases. More advanced techniques like immunohistochemistry use antibodies to identify proteins, while in situ hybridization identifies DNA and RNA, assisting in diagnosis of the remaining cases. Together these staining methods are essential for pathologists to examine tissue at a microscopic level and determine if a sample is benign or malignant.

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    © All Rights Reserved
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    19 views9 pages

    Stain Techniques

    Uploaded by

    rafael santamaría
    Staining techniques are used to diagnose tissue samples under the microscope. The main techniques are hematoxylin and eosin staining, special stains, immunohistochemistry, and in situ hybridization. Hematoxylin and eosin staining uses dyes to differentiate cell structures and can provide a final diagnosis in 70-80% of cases. More advanced techniques like immunohistochemistry use antibodies to identify proteins, while in situ hybridization identifies DNA and RNA, assisting in diagnosis of the remaining cases. Together these staining methods are essential for pathologists to examine tissue at a microscopic level and determine if a sample is benign or malignant.

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    © All Rights Reserved
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    of 9

    Introduction to staining techniques

    Which technique for which diagnosis?

    Image
    Here
    Understand the word "staining"
    The staining step

    + microscope

    STAINING DIAGNOSIS
    STEP

    +pathologist
    Tissue section on a Stained slide =
    glass slide stained tissue
    section on a glass
    slide
    Understand the main staining techniques
    Which techniques?
    Tissue section on
    a glass slide

    100%
    Cell shape
    H&E 70-80%
    Hematoxylin and Eosin
    final diagnostic
    Malignant Benign or Malignant

    10 %
    SpSt 15 % 3-4 % ISH
    Special Stains In Situ Hybridization

    Cell & tissue shape & chemistry IHC DNA & RNA
    Immuno Histo Chemistry

    Proteins -
    Antigens
    Understand the main staining techniques
    Techniques classification
    • Primary staining
    – Dye based
    • Hematoxylin & Eosin ( H&E )
    • Special Stains
    • Advanced staining
    – More complex, more steps
    • Immunohistochemistry ( IHC )
    • Fluorescence Antibodies ( FITC )
    • In situ hybridisation ( ISH )
    – Silver ISH ( SISH )
    – Chromogenic ISH ( CISH )
    – Fluorescent ISH ( FISH )
    Understand the staining techniques
    Hematoxylin & Eosin
    • Primary staining - Dye based – Hematoxylin and
    Eosin

    Tissue(/cells) on a glass slide

    + Hem
    Eosi
    n
    TARGET = nuclei for Hematoxyline, cytoplasm and stroma for Eosin

    After review by a pathologist: final diagnosis (70-80% cases)


    Understand the staining techniques
    Special Stains

    • Primary staining - Dye based – Many different dies


    (color)
    Jones light green -
    Kidney

    + Dye cocktail

    Tissue(/cells) on a glass slide Elastic -


    Artery
    TARGET = Protein, Lipids, Sugar, nucleic acids depending on the dye used

    H&E result + SpSt result reviewed by a pathologist: final


    diagnosis
    Trichr. II blue -
    Liver
    Understand the staining techniques
    IHC
    • Advanced staining – Immuno Histo Chemistry - Antibody
    based

    Her2neu on
    + + breast

    Tissue(/cells) on a glass slide Antibody Detection

    TARGET = ANTIGEN (Part of PROTEIN)

    H&E result + IHC result reviewed by a pathologist: final CD3 on tonsil


    diagnosis
    Understand the staining techniques
    SISH
    ISH
    • Advanced staining – In Situ Hybridization - Probe
    based
    SISH on
    breast
    CISH

    + +
    HPV on cervix
    Tissue(/cells) on a glass slide Nucleic Acid Probe Detection
    FISH
    TARGET = Nucleic Acids (DNA, RNA)
    (to be discontinued)

    H&E + IHC + ISH result reviewed by a pathologist: final


    diagnosis
    FISH on
    Understand the staining techniques
    Summary
    H&E - Hematoxylin and Eosin
    Two-color differentiation of various structures related to
    size, shape and arrangement B/W aerial
    Used as screening test. Final diagnosis possible in photo
    70-80% of cases
    Primary staining

    SS – Special Stains
    Uses various dyes, methods and chemical procedures. Color aerial
    Further identifies broad structures and cellular processes photo
    (10% of cases)
    Primary staining

    IHC – ImmunoHistoChemistry
    Antibodies used to selectively identify cellular products
    View factory
    (proteins). 15% cases.

    Advanced staining

    ISH – In Situ Hybridization


    DNA and RNA: Used to look for genes or gene
    expression. 3 – 4% cases.
    Blueprint
    Advanced staining

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