MTAP 2 - Infiltration to Embedding Quiz
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Questions and Answers

What is a primary limitation of using heat-based paraffin wax infiltration for tissue processing?

  • It is not suitable for tissues with high fat content. (correct)
  • It requires excessive time for complete infiltration.
  • It causes significant tissue shrinkage.
  • It is ineffective for cytology specimens.
  • Which of the following is NOT typically part of the initial steps of tissue processing?

  • Staining (correct)
  • Dehydration
  • Fixation
  • Clearing
  • What is the main purpose of using ascending concentrations of ethyl alcohol in tissue processing?

  • To remove water from the tissue. (correct)
  • To dissolve any remaining fats in the tissue.
  • To harden the tissue for cutting.
  • To stain the tissue for visualization.
  • What is the typical temperature range for an oven used in manual paraffin wax infiltration?

    <p>55-60 degrees Celsius. (D)</p> Signup and view all the answers

    Which of the following is the most common clearing agent used in tissue processing?

    <p>Xylene (C)</p> Signup and view all the answers

    What is a key advantage of using Carbowax in tissue processing?

    <p>It is water-soluble which eliminates the need for clearing and dehydration. (D)</p> Signup and view all the answers

    Which embedding medium is specially suitable for producing sections of eye specimens?

    <p>Bioloid or Embeddol (A)</p> Signup and view all the answers

    Which characteristic of Ester wax differentiates it from paraffin wax?

    <p>It has a lower melting point but produces harder sections. (D)</p> Signup and view all the answers

    What is the purpose of the 'embedding' process in tissue preparation?

    <p>To produce a tissue block that can be used for sectioning. (D)</p> Signup and view all the answers

    What primary advantage does negative atmospheric pressure offer during the infiltration step?

    <p>It ensures rapid removal of bubbles, in addition to the clearing agent. (C)</p> Signup and view all the answers

    What is the primary purpose of infiltration in tissue processing?

    <p>To fill cavities and tissue spaces. (D)</p> Signup and view all the answers

    Why is a firm consistency essential for tissue specimens after infiltration?

    <p>To facilitate easier and more precise cutting. (D)</p> Signup and view all the answers

    Which of the following is the most commonly used medium for infiltration?

    <p>Paraffin wax. (D)</p> Signup and view all the answers

    At what temperature does the paraffin wax used for infiltration typically melt?

    <p>56 degrees C. (C)</p> Signup and view all the answers

    What does constant agitation accomplish in the infiltration process?

    <p>It promotes rapid entry of paraffin wax into the tissue. (D)</p> Signup and view all the answers

    What is the primary function of the autotechnicon in tissue processing?

    <p>To automatically carry out the first few steps including infiltration. (C)</p> Signup and view all the answers

    Which of the following methods uses negative atmospheric pressure inside an oven to speed up the paraffin wax infiltration?

    <p>Vacuum-assisted paraffin wax infiltration. (B)</p> Signup and view all the answers

    How many changes of wax are typically needed during infiltration to ensure complete impregnation?

    <p>2-3 Changes (C)</p> Signup and view all the answers

    What is the primary purpose of using a float-out bath in histological procedures?

    <p>To flatten the tissue ribbons and remove wrinkles. (D)</p> Signup and view all the answers

    Which material is LEAST advisable for stropping a knife, according to the text?

    <p>Mineral oil coated leather (B)</p> Signup and view all the answers

    What is the recommended temperature range for a float out bath in degrees Celsius?

    <p>45-50 (B)</p> Signup and view all the answers

    What type of movement should be used when stropping a knife?

    <p>Toe to heel movement, from base to tip (C)</p> Signup and view all the answers

    What is the process of 'fishing out' in the context of histology?

    <p>Removing ribbons from the float out bath (D)</p> Signup and view all the answers

    Which material is most commonly utilized as an embedding medium?

    <p>Paraffin wax (A)</p> Signup and view all the answers

    What is the recommended temperature for rapid cooling of paraffin during embedding?

    <p>-5°C (C)</p> Signup and view all the answers

    What is a key advantage of using plastic embedding rings, such as Tissue-Tek?

    <p>They minimize the amount of paraffin wax required. (C)</p> Signup and view all the answers

    Which substance should be used to coat the mold for easy removal of the paraffin block?

    <p>Glycerin (C)</p> Signup and view all the answers

    What is the main purpose of using a compound embedding unit?

    <p>To embed several specimens simultaneously (D)</p> Signup and view all the answers

    Which of these options is NOT a type of mold mentioned for embedding?

    <p>Glass slides (D)</p> Signup and view all the answers

    What does the term 'infiltration' refer to in the context of embedding?

    <p>The process of replacing water with an embedding medium (C)</p> Signup and view all the answers

    What is the primary function of formalin when used in gelatin infiltration?

    <p>To harden the tissue before sectioning (C)</p> Signup and view all the answers

    Why is the orientation of the specimen considered easy when using plastic embedding rings or Bass molds?

    <p>Because they allow visual placement of the tissue, and have uniform cavities (B)</p> Signup and view all the answers

    Besides a tissue softener in decalcification, what is another use for phenol?

    <p>As an antimicrobial agent to prevent mold growth (A)</p> Signup and view all the answers

    What is the main purpose of the embedding process in tissue preparation?

    <p>To place the tissue in a medium for cutting/sectioning (D)</p> Signup and view all the answers

    Which of the following describes the standard solution and time required for tissue immersion?

    <p>10% formalin for 12-24 hours (C)</p> Signup and view all the answers

    What are the common embedding mediums used?

    <p>Paraffin wax and plastic resin (A)</p> Signup and view all the answers

    What characterizes Leuchhart's embedding molds?

    <p>They are L-shaped molds made of heavy brass or metal (C)</p> Signup and view all the answers

    What is the alternative name for 'embedding' in tissue processing?

    <p>Casting/Blocking (C)</p> Signup and view all the answers

    What is an advantage of using Leuchhart’s embedding molds that is mentioned in the content?

    <p>You can produce tissue blocks of different sizes (C)</p> Signup and view all the answers

    Flashcards

    Fixation

    The initial step in tissue processing that uses formaldehye to preserve cell structures and harden the tissue for easier cutting.

    Dehydration

    The process of gradually removing water from the tissue using increasing concentrations of alcohol, preparing it for paraffin wax.

    Clearing

    The process of removing alcohol from the tissue using a clearing agent like xylene, making the tissue transparent for wax infiltration.

    Infiltration

    The process of replacing the clearing agent with molten paraffin wax, allowing the tissue to become embedded in the wax for cutting.

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    Paraffin wax infiltration

    A method of embedding tissue in paraffin wax that involves placing the tissue in a wax bath at a specific temperature and changing the wax at intervals.

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    Paraffin Wax

    A medium used for infiltration that turns solid at room temperature, making it easier to cut tissue sections.

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    Melting Point

    The temperature at which a substance changes from solid to liquid. For paraffin wax, it's 56 degrees Celsius.

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    Autotechnicon

    An automated machine used for processing tissues. It automatically completes the first 4 steps including fixation, dehydration clearing, and infiltration.

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    Heat Assisted Infiltration

    Using heat to speed up the process of filling tissue spaces with paraffin wax.

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    Vacuum Infiltration

    Creating a negative pressure inside the oven to accelerate the penetration of paraffin wax into tissue spaces.

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    Agitation

    The process of moving the tissue around within the wax solution to ensure that all areas are fully infiltrated.

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    Multiple Changes of Wax

    Repeated changes of fresh paraffin wax to ensure complete removal of the clearing agent and thorough penetration of the wax into the tissue.

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    Tissue Embedding

    This method involves replacing water in tissue with a supporting medium like paraffin wax, allowing for thin tissue sectioning.

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    Ester Wax

    A type of wax used for embedding, typically with a lower melting point but harder consistency.

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    Paraplast

    Recommended for bones and brain specimens due to its suitability for harder tissues.

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    Carbowax

    A water-soluble wax, eliminating the need for clearing and dehydration steps. Useful for enzyme studies.

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    Celloidin Infiltration

    A method of embedding tissues using celloidin, a type of nitrocellulose, as the supporting medium.

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    Stropping

    A specialized process where a leather strop is used to polish and refine the sharp edge of a knife, removing burrs and irregularities after honing.

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    Float out bath

    The process of flattening ribbons of tissue by removing creases and wrinkles using a float out bath.

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    Fishing out

    The act of carefully removing the tissue ribbons from the float out bath without disturbing their structure.

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    Orientation

    The specific placement of tissue ribbons in the centre of a slide, ensuring proper alignment for further processing.

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    Mineral Oil

    A type of oil that should not be used on leather strops because it can damage the leather, causing blisters or deterioration.

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    Embedding

    The process of embedding a tissue sample in a mold filled with a solid medium, like paraffin wax or resin, to create a block for sectioning.

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    Embedding medium

    A solid material, like paraffin wax or plastic resin, that is used to solidify around a tissue sample during embedding.

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    Embedding mold

    A container used to hold the tissue sample and embedding medium during the embedding process.

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    Leuchhart’s embedding molds

    A specific type of embedding mold made with two L-shaped parts that can be fitted together to hold a tissue sample.

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    Phenol in embedding

    A commonly used chemical in embedding solutions to prevent the growth of mold and maintain the quality of the tissue blocks.

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    Gelatin embedding

    A type of embedding medium that uses gelatin to solidify around the tissue sample, providing a pliable and flexible block for sectioning.

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    Formalin in gelatin embedding

    The purpose of formalin in gelatin embedding is to harden the tissue before cutting. It is not used as a fixative.

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    Casting/Blocking

    The process of embedding can be referred to as casting or blocking, as it results in creating a tissue block that is ready for sectioning.

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    Manual embedding

    A manual method of embedding where tissue is placed in a mold filled with melted paraffin, then cooled rapidly, allowing for quick and easy orientation.

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    Interlocking plate mold

    A type of mold that consists of interconnected plates, allowing the user to create multiple blocks of tissue simultaneously.

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    Tissue-Tek plastic embedding rings

    A plastic embedding ring designed specifically for embedding tissues in paraffin wax; commonly used with Bass molds.

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    Bass molds

    A specific type of embedding mold that incorporates a base and removable plates, providing a convenient way to create multiple blocks of tissue.

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    Paper boats

    A tool used for embedding tissues using paraffin wax, allowing for easy removal of the block once it solidifies.

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    Study Notes

    MTAP 2 - Infiltration to Embedding

    • Histopathology specimens are processed similarly, regardless of type (autopsy, biopsy, surgical, cytology)
    • Fixation preserves tissues to harden them for cutting
    • Dehydration uses ascending ethanol concentrations
    • Clearing uses xylene as a common agent
    • Infiltration (impregnation) fills tissue spaces for consistent firmness for cutting
    • Paraffin wax infiltration is the most common method, using an oven for a consistent temperature

    Disadvantages of Paraffin Wax Infiltration

    • Not suitable for fatty tissues due to melting
    • Requires longer time for dense specimens

    Methods of Paraffin Wax Infiltration

    • Manual: Uses a dissecting pan with melted paraffin wax, placing the tissue in the pan and into an oven set 2–5°C above the wax's melting point. The process involves four 15-minute intervals over an hour.
    • Automatic (Autotechnicon): An automatic tissue processor performs fixation, dehydration, clearing, and infiltration
    • Requires less paraffin wax
    • Rapid embedding
    • Easy specimen orientation

    Substitutes for Paraffin Wax

    • Paraplast: For bones, brain specimens
    • Bioloid/Embeddol: For eye specimens
    • Carbowax: Water-soluble, eliminates clearing and dehydration, suitable for enzyme studies

    Celloidin Infiltration

    • No heat required; slow method
    • Appropriate for specimens with significant cavities
    • Requires 3 containers of increasing celloidin concentrations (2-4%, 4-6%, and 8-12%), immersing the specimens for varying durations

    Gelatin Infiltration

    • Recommended for eye specimens and enzyme studies
    • Immerse the specimen in a gelatin solution for 24 hours (10% Gelatin + 1% phenol), then 12 hours (20% Gelatin + 1% phenol))

    Embedding

    • Places tissue into a mold with embedding medium.
    • Paraffin wax is the most common embedding medium
    • Produces tissue blocks suitable for cutting

    Embedding Molds

    • Disposable and non-disposable molds
    • Hulmahan embedding molds
    • Leuchhart's embedding molds (“L” pieces)
    • Compound embedding unit

    Types of Microtomes

    • Ultrathin Microtome: For electron microscopy (0.5 µm thickness)
    • Rotary Microtome: For paraffin embedded tissues (4-6 µm thickness)
    • Rocking Microtome: (10-12 µm thickness)
    • Sliding Microtome: (7–9 µm thickness)
    • Base Sledge Microtome: Movable part is the block holder
    • Cryostat: For immediately hardening fresh tissue using temperature
    • Freezing Microtome: For tissues with heat-sensitive structures
    • Diamond Knife: For electron microscopy

    Types of Blades

    • Biconcave: For paraffin tissue
    • Plane Concave: Cut tough samples in paraffin

    Honing and Stropping

    • Honing: Removes nicks and irregularities from the blade
    • Stropping: Polishes and sharpens the blade further

    Paraffin Removal (Deparaffinization)

    • Removes paraffin and clearing agents to ready samples for staining

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    Description

    Test your knowledge on the processes of histopathology specimens, focusing on paraffin wax infiltration methods and their advantages and disadvantages. This quiz will cover fixation, dehydration, clearing, and the techniques used in embedding tissues for microscopic evaluation.

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