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    Clearing and Infiltration (Histopathology and Cytology)

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    Noriz Ember Dominguez
    1. If tissue is incompletely dehydrated, the clearing agent will not perform its function properly and the tissue will not be prepared for embedding, resulting in a soft, mushy tissue block. 2. Xylene is the best clearing agent for routine work because it clears tissue quickly, is miscible with alcohol and paraffin, and does not extract dyes, allowing for effective clearing and replacement by wax during embedding. 3. Factors that affect clearing include the clearing agent's boiling point, viscosity, and prolonged exposure, which can cause tissue brittleness and difficult sectioning.

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    Clearing and Infiltration (Histopathology and Cytology)

    Uploaded by

    Noriz Ember Dominguez
    56 views2 pages
    1. If tissue is incompletely dehydrated, the clearing agent will not perform its function properly and the tissue will not be prepared for embedding, resulting in a soft, mushy tissue block. 2. Xylene is the best clearing agent for routine work because it clears tissue quickly, is miscible with alcohol and paraffin, and does not extract dyes, allowing for effective clearing and replacement by wax during embedding. 3. Factors that affect clearing include the clearing agent's boiling point, viscosity, and prolonged exposure, which can cause tissue brittleness and difficult sectioning.

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    CLEARING AND INFILTRATION (HISTOPATHOLOGY AND CYTOLOGY)

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    1. If tissue is incompletely dehydrated, the clearing agent will not perform its function properly and the tissue will not be prepared for embedding, resulting in a soft, mushy tissue block. 2. Xylene is the best clearing agent for routine work because it clears tissue quickly, is miscible with alcohol and paraffin, and does not extract dyes, allowing for effective clearing and replacement by wax during embedding. 3. Factors that affect clearing include the clearing agent's boiling point, viscosity, and prolonged exposure, which can cause tissue brittleness and difficult sectioning.

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    © All Rights Reserved
    Download as PDF, TXT or read online from Scribd
    Download as pdf or txt
    56 views2 pages

    Clearing and Infiltration (Histopathology and Cytology)

    Uploaded by

    Noriz Ember Dominguez
    1. If tissue is incompletely dehydrated, the clearing agent will not perform its function properly and the tissue will not be prepared for embedding, resulting in a soft, mushy tissue block. 2. Xylene is the best clearing agent for routine work because it clears tissue quickly, is miscible with alcohol and paraffin, and does not extract dyes, allowing for effective clearing and replacement by wax during embedding. 3. Factors that affect clearing include the clearing agent's boiling point, viscosity, and prolonged exposure, which can cause tissue brittleness and difficult sectioning.

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    © All Rights Reserved
    Download as PDF, TXT or read online from Scribd
    Download as pdf or txt
    of 2

    1. What will happen to the clearing agent if the tissue is incompletely dehydrated?

    Complete dehydration should be achieved before proceeding to the next


    routine processing, which is clearing or dealcoholization since most clearing agents are
    not miscible with aqueous fixatives. Clearing, on the other hand, is performed to remove
    alcohol or the dehydrating agent from the tissue and replace it with a clearing agent
    prior to impregnation and embedding since alcohol is highly immiscible with wax.
    However, if the tissue is not completely dehydrated, clearing agents such as xylene will
    not act accordingly or perform its function since the tissue may still contain the aqueous
    fixative and water in which the clearing agent is immiscible with. As a result of
    inadequate dehydration, the tissue will not be prepared prior to impregnation resulting
    in a soft, mushy tissue block.

    2. What is the best clearing agent for routine work and why?

    Xylene is the best clearing agent for routine work because it has the different
    characteristics of a good clearing agent. It clears within 15 to 30 minutes and makes
    tissues transparent. It is miscible with absolute alcohol as well as paraffin, and it does not
    extract out aniline dyes. Xylene is readily replaced by wax during impregnation and
    embedding since it evaporates quickly in a paraffin oven. Also, it is reasonably cost
    effective and works well for short-term clearing of tissue blocks.

    3. What are the factors affecting clearing of specimens?

    There are several factors that affect the clearing or dealcoholization of


    specimens including the clearing agent’s boiling point, viscosity, as well as prolonged
    exposure to the clearing agent. Clearing agents that have low boiling point are usually
    more easily replaced by melted paraffin. Viscosity, on the hand, affects the speed of
    penetration of the clearing agent. In addition, prolonged exposure to most clearing
    agents may lead to brittleness of the tissue, thereby making it more difficult to cut.

    4. What are the precautionary/safety measures to be observed in performing paraffin


    infiltration?

    The following are some of precautionary/safety measures that must be taken into
    consideration when performing paraffin infiltration: (1) the tissue should not be left in the
    paraffin over for longer than 4 hours; (2) do not overheat the tissue to avoid hardening
    and brittleness; (3) the paraffin oven should be maintained at a temperature 2- to
    5-degree Celsius above the melting point of the paraffin; (4) paraffin wax should be free
    from dust, water droplets, and other other foreign matter; (5) before using a fresh wax, it
    must be filtered in a wax oven at a temperature 2-degree Celsius higher than its melting
    point; (6) wax that has been trimmed away from the infiltrated tissue may be melted
    and filtered for future use; (7) paraffin wax may be used only twice.

    5. What is the effect of excessively heating the paraffin more than the recommended
    temperature?

    Excessively heating the paraffin more than the recommended temperature will
    have the same effect as prolonged treatment in melted paraffin. Tissues become
    increasingly harder and more brittle as they are heated. Therefore, overheated paraffin
    will cause shrinkage and hardening of the tissues, and destroy lymphoid tissues
    completely. Due to this reason, the paraffin oven should be maintained at a
    recommended temperature of 2- to 5-degree Celsius above the melting point of
    paraffin to be used in infiltration.

    6. What is the importance of maintaining the temperature of the paraffin oven at 2-3
    degrees celsius higher than the melting point of the paraffin wax?

    The proper temperature of the paraffin is critical during infiltration. If the tissue is
    exposed to overheated paraffin, it will cause the tissue to be brittle. However, if the
    paraffin is below the recommended temperature range, infiltration of the tissue may not
    be successful. Therefore, it is important that the temperature of the paraffin oven is
    maintained at 2- to 3-degree Celsius higher than the melting point of the paraffin wax to
    prevent relative shrinkage and hardening of the tissue. Also, maintaining the
    temperature of the paraffin oven at the said range will allow proper and complete
    infiltration of the tissue.

    References:

    Jocelyn H. Bruce-Gregorios. (2016.). Histopathologic Techniques. Katha Publishing Co.


    Temperature Factor—LabCE.com, Laboratory Continuing Education. (n.d.). Retrieved
    November 25, 2021, from https://www.labce.com/spg572631_temperature_factor.aspx
    Tissue Clearing—LabCE.com, Laboratory Continuing Education. (n.d.). Retrieved November 25,
    2021, from https://www.labce.com/spg572642_tissue_clearing.aspx

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