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  • Artifactual pigments and their removal methods

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    Glenn Perez
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    This document describes various artifactural pigments that can be introduced during tissue fixation and staining, along with methods for their removal. It lists the appearance and solvents that can remove precipitates from paraformaldehyde, acid formaldehyde hematin, mercuric chloride, chromate fixatives, osmic oxide, and malarial pigments. Common washing out processes and types of fixatives used are also outlined.

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    Histopath Must Know

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    This document describes various artifactural pigments that can be introduced during tissue fixation and staining, along with methods for their removal. It lists the appearance and solvents that can remove precipitates from paraformaldehyde, acid formaldehyde hematin, mercuric chloride, chromate fixatives, osmic oxide, and malarial pigments. Common washing out processes and types of fixatives used are also outlined.

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    © All Rights Reserved
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    19 views2 pages

    Artifactual pigments and their removal methods

    Uploaded by

    Glenn Perez
    This document describes various artifactural pigments that can be introduced during tissue fixation and staining, along with methods for their removal. It lists the appearance and solvents that can remove precipitates from paraformaldehyde, acid formaldehyde hematin, mercuric chloride, chromate fixatives, osmic oxide, and malarial pigments. Common washing out processes and types of fixatives used are also outlined.

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    of 2

    ARTIFACTUAL PIGMENTS

    PIGMENT APPEARANCE REMOVED BY


    Paraformaldehyde White crystalline precipitate 1. 10% methanol
    2. Filtration methods
    Acid Formaldehyde Hematin Brown or black granules 1. 1% alcoholic KOH
    (Formic acid + Hgb) - May obscure 2. Saturated picric acid
    microscopic details 3. Kardasewitch
    method (ammonia
    water + ethanol)
    4. Lillie’s method
    (ammonia water +
    alcohol +acetone +
    hydrogen peroxide)
    Mercuric chloride Black granular deposits Alcoholic iodine
    (saturated iodine
    solution in 96%
    alcohol)
    DE-ZENKERIZATION
    Chromate fixatives Fine yellow or brown deposits Acid alcohol
    Osmic oxide (Osmium Black precipitate Cold water
    tetroxide
    Malarial pigments Brown black granules Saturated alcoholic
    (Hemozoin) (intracellular) picric acid
    - Usually mistaken as
    Acid formaldehyde
    hematin
    WASHING OUT – process of removing excess fixative from tissue
    1. Tap water – chromates, formalin and osmic acid
    2. 50-70% alcohol – picric acid
    3. Alcoholic iodine – mercuric chloride
    FIXATIVES
    1. ALDEHYDE
    2. METALLIC
    a. Mercuric chloride (5-7%)
    b. Chromate
    c. Lead
    3. PICRIC ACID
    4. GLACIAL ACETIC ACID
    5. ALCOHOL
    6. OSMIUM TETROXIDE (OSMIC ACID)
    7. TRICHOLORACTERIC ACID
    8. ACETONE
    9. HEAT

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