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    Clearing &amp Dealcoholization

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    bubblyeivinej
    Clearing, or dealcoholization, is the process of removing alcohol from tissue sections by immersing them in a clearing agent to make the tissue transparent. Common clearing agents include benzene, chloroform, xylene, toluene, cedar oil, clove oil, and gasoline. Each agent has advantages and disadvantages in terms of clearing time, shrinkage effects, toxicity, and other properties. Clearing makes tissue transparent and prepares it for further processing like wax embedding and mounting of stained sections.

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    Clearing &amp Dealcoholization

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    bubblyeivinej
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    Clearing, or dealcoholization, is the process of removing alcohol from tissue sections by immersing them in a clearing agent to make the tissue transparent. Common clearing agents include benzene, chloroform, xylene, toluene, cedar oil, clove oil, and gasoline. Each agent has advantages and disadvantages in terms of clearing time, shrinkage effects, toxicity, and other properties. Clearing makes tissue transparent and prepares it for further processing like wax embedding and mounting of stained sections.

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    Clearing & Dealcoholization

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    Clearing, or dealcoholization, is the process of removing alcohol from tissue sections by immersing them in a clearing agent to make the tissue transparent. Common clearing agents include benzene, chloroform, xylene, toluene, cedar oil, clove oil, and gasoline. Each agent has advantages and disadvantages in terms of clearing time, shrinkage effects, toxicity, and other properties. Clearing makes tissue transparent and prepares it for further processing like wax embedding and mounting of stained sections.

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    Download as ppt, pdf, or txt
    2K views23 pages

    Clearing &amp Dealcoholization

    Uploaded by

    bubblyeivinej
    Clearing, or dealcoholization, is the process of removing alcohol from tissue sections by immersing them in a clearing agent to make the tissue transparent. Common clearing agents include benzene, chloroform, xylene, toluene, cedar oil, clove oil, and gasoline. Each agent has advantages and disadvantages in terms of clearing time, shrinkage effects, toxicity, and other properties. Clearing makes tissue transparent and prepares it for further processing like wax embedding and mounting of stained sections.

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    HISTOPATHOLOGY

    CLEARING & DEALCOHOLIZATION


    Jenivie Morillo Jose University of Santo Tomas National Childrens Hospital

    DEFINITION:
    CLEARING / DEALCOHOLIZATION Process of removing alcohol from section of tissue by immersing them in ANTE MEDIUM Tissue becomes transparent as a result of ante medium raising the REFRACTIVE INDEX

    MOST COMMON CLEARING AGENTS


    1. BENZENE CLEARING TIME: 15-60 minutes
    BEST ROUTINE CLEARING AGENT for urgent biopsies Rapid in action HIGHLY INFLAMMABLE, CARCINOGENIC

    IDEAL CHARACTERISTICS:
    Miscible with alcohol Miscible with paraffin wax Should not produce excessive tissue shrinkage and hardening Should not evaporate quickly in the water bath

    Should make tissue transparent

    MOST COMMON CLEARING AGENTS


    2. CHLOROFORM CLEARING TIME: 6- 24 HOURS ONE BATH is needed Miscible with ABSOLUTE ALCOHOL Causes very little SHRINKAGE and hardening of tissues Suitable for LARGE TISSUE SPECIMENS, TOUGH TISSUES(skin, fibroid, decalcified tissues), for NERVOUS TISSUE, LYMPH NODES and EMBRYOS Slowly DISPLACES alcohol TOXIC to the LIVER

    MOST COMMON CLEARING AGENTS


    3. XYLENE(XYLOL)
    CLEARING TIME: 15-30 minutes ONE BATH is needed Miscible with ABSOLUTE ALCOHOL COLORLESS

    For URGENT BIOPSIES


    Rapid in action Makes tissue TRANSPARENT

    MOST COMMON CLEARING AGENTS


    3. XYLENE(XYLOL) *continuation*
    NOT CARCINOGENIC Cheap HIGHLY INFLAMMABLE VOLATILIZED rapidly in PARAFFIN OVEN

    EXCESSIVE tissue shrinkage and


    hardening MILKY if INCOMPLETELY DEHYDRATED

    MOST COMMON CLEARING AGENTS


    4. TOULENE
    CLEARING TIME: 15-60 minutes ONE BATH is needed Miscible with ABSOLUTE ALCOHOL Rapid in action

    NOT CARCINOGENIC
    HIGHLY INFLAMMABLE Shrinkage of tissues

    MOST COMMON CLEARING AGENTS


    5. CEDAR OIL For CENTRAL NERVOUS SYSTEM, SKIN and SMOOTH MUSCLE and CYTOLOGICAL WORK CLEARING TIME: Indefinite until clear TWO BATHS Miscible with 95% alcohol BEST CLEARING AGENT Minimal shrinkage and hardening of the tissues Extremely slow Expensive

    MOST COMMON CLEARING AGENTS


    6. CLOVE OIL
    For SKIN and SMOOTH MUSCLE CLEARING TIME: Indefinite TWO BATHS Miscible with 95% alcohol

    LITTLE shrinkage
    Wax impregnation is SLOW and DIFFICULT Makes tissue BRITTLE

    MOST COMMON CLEARING AGENTS


    7. OIL OF BERGAMOT
    For SKIN and SMOOTH MUSCLE CLEARING TIME: Indefinite TWO BATHS Miscible with 95% alcohol

    NOT EXTRACT aniline dyes


    Used with CELLOIDIN PREPARATION Expensive

    MOST COMMON CLEARING AGENTS


    8. OIL OF ORIGANUM
    Spanish hop oil or thyme oil or mixtures of these two, with or without turpentine For SKIN CLEARING TIME: Indefinite TWO BATHS miscible with 95% alcohol DOESNT DISSOLVE celloidin DISSOLVES ANILINE DYES Expensive

    MOST COMMON CLEARING AGENTS


    9. OIL OF WINTERGREEN
    methyl salicylate; artificial oil For DELICATE TISSUES CLEARING TIME: Indefinite TWO BATHS

    Miscible with 95% alcohol

    MOST COMMON CLEARING AGENTS


    9. OIL OF WINTERGREEN *continuation*
    HARDENING EFFECT OF ALCOHOL can

    be avoided
    Mainly for CELLOIDIN SECTIONS but must be soaked by CHLOROFORM or XYLENE before mounting POOR DEHYDRANT OXIDIZES easily on STANDING

    MOST COMMON CLEARING AGENTS


    10. CARBON DISULFIDE For SMOOTH MUSCLE
    CLEARING TIME: Indefinite TWO BATHS Minimal shrinkage of tissue VERY INFLAMMABLE, with FOUL ODOR

    Wax impregnation is SLOW and DIFFICULT

    MOST COMMON CLEARING AGENTS


    11. CARBON TETRACHLORIDE

    For ROUTINE work Similar to CHLOROFORM

    MOST COMMON CLEARING AGENTS


    12. CARBOL XYLENE For FRIABLE TISSUES
    CLEARING TIME: up to 1 hour Made by adding ANHYDROUS CRYSTALS of PHENOL to XYLENE until no more dissolves

    Good for MOIST and HUMID CLIMATES


    CORROSIVE

    MOST COMMON CLEARING AGENTS


    13. TETRAHYDROFURAN For CELLULAR ORGANS, FRIABLE TISSUES, SKIN, ATHERMATOUS ARTERIES, HARD COLLAGENOUS SPECIMENS CLEARING TIME: 2-4 hours THREE BATHS Colorless liquid with 65C (boiling point), -108.5C (freezing point) and 0.8894 (density) Wide range solvent (including fats) Soluble in most solvents (lower alcohols, ketones, esters, chloroform, ether, acetone, benzene, zylene, dioxane, creosote)

    MOST COMMON CLEARING AGENTS


    13. TETRAHYDROFURAN *continuation* SUPERIOR to ORDINARY DEHYDRATING CLEARING AGENTS (can perform 2 processes at the same time) SHORTENS the TOTAL PROCESSING TIME and allowing MORE time for fixation

    Causes CONJUNCTIVAL IRRITATION for about 6


    MONTHS OFFENSIVE ODOR

    MOST COMMON CLEARING AGENTS


    14. TERPINEOL EXCELLENT for DELICATE MATERIAL

    especially EYES
    CLEARING TIME: 2-4 hours TWO BATHS

    Miscible with 95% alcohol


    Solution consists of 1 part XYLENE and 4 parts TERPINEOL Expensive

    MOST COMMON CLEARING AGENTS


    15. PHENOL For SMOOTH MUSCLE
    CLEARING TIME: 2 hours TWO BATHS Good for PARAFFIN and CELLOIDIN WORK

    DEHYDRATES and CLEARS very quickly


    VOLATILE

    MOST COMMON CLEARING AGENTS


    16. HIGH TESTAVIATION LEAD FREE GASOLINE
    EXCELLENT CLEARING AND DEALCOHOLIZATION AGENT Cedar oil, gasoline and petroleum ether cause the LEAST HARDENING of tissues

    APPLICATIONS OF CLEARING IN HISTOPATHOLOGY:


    1. To make tissue, embryos and parasite

    transparent
    2. Dealcoholization of tissue preparatory to wax impregnation 3. Dealcoholization of stained sections prior to mounting in PERMOUNT, CLARITE or CANADA BALSAM

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