Light and Electron Microscopy Overview
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Questions and Answers

What is the resolution power of an electron microscope?

  • 10 nm
  • 1 micron
  • 0.2 micron
  • 0.2 nm (correct)
  • Which stain is most suitable for visualizing lipids?

  • Hematoxylin and eosin stain
  • Sudan stain (correct)
  • Masson's trichrome stain
  • Silver impregnation
  • What is the primary purpose of fixation in fresh tissue?

  • Prevent putrefaction and autolysis (correct)
  • Clear the sections
  • Remove water
  • Replace alcohol
  • Which of the following statements about Hematoxylin is accurate?

    <p>It is a basic dye</p> Signup and view all the answers

    What is the definition of vital stain?

    <p>Staining of a living cell inside the body</p> Signup and view all the answers

    What is the standard contrasting technique used in transmission electron microscopy?

    <p>Uranyl acetate and lead citrate</p> Signup and view all the answers

    What color does glycogen stain using Best's carmine stain?

    <p>Red</p> Signup and view all the answers

    Which method is used to detect ATPase enzyme in muscle fibers?

    <p>Enzyme histochemistry</p> Signup and view all the answers

    What type of microscope utilizes fluorescent light?

    <p>Fluorescent microscope</p> Signup and view all the answers

    What is the principle behind immunohistochemistry?

    <p>Reaction between antibody and antigen</p> Signup and view all the answers

    In histochemistry, what color is observed when detecting nucleic acids using the Feulgen reaction?

    <p>Red to purple</p> Signup and view all the answers

    Which staining method is used to visualize lipids in tissue?

    <p>Sudan III</p> Signup and view all the answers

    What type of fixation method is used in electron microscopy to preserve ultrastructural details?

    <p>Double fixation with glutaraldehyde and osmium tetraoxide</p> Signup and view all the answers

    What is the primary purpose of eosin in histological staining?

    <p>To provide a pink color to acidophilic cytoplasm</p> Signup and view all the answers

    Which of the following statements about neutral stains is correct?

    <p>They combine basic and acidic dyes to stain all components.</p> Signup and view all the answers

    Which statement accurately describes vital staining?

    <p>It involves injecting a non-toxic dye into a living animal.</p> Signup and view all the answers

    What color does Masson’s stain give to collagen?

    <p>Green</p> Signup and view all the answers

    What phenomenon results in a metachromatic stain?

    <p>Generation of a color different from the dye used</p> Signup and view all the answers

    Which of the following dyes is commonly used in supravital staining?

    <p>Methylene blue</p> Signup and view all the answers

    How do supravital stains primarily differ from vital stains?

    <p>Supravital stains are used in vitro, whereas vital stains are in vivo.</p> Signup and view all the answers

    Which type of stain is known to stain specific cell components with a color change?

    <p>Metachromatic stain</p> Signup and view all the answers

    What is the purpose of deparaffinization in the paraffin technique?

    <p>To remove paraffin for easy staining</p> Signup and view all the answers

    Which staining technique is primarily used for observing nuclear structures?

    <p>Hematoxylin staining</p> Signup and view all the answers

    What is the correct order of steps involved in staining paraffin sections?

    <p>Deparaffinization, Staining, Hydration, Dehydration</p> Signup and view all the answers

    What charge do acidic stains carry, and what do they typically stain?

    <p>Negative charge; acidic structures</p> Signup and view all the answers

    During the hydration step, which sequence of alcohol concentrations is used?

    <p>100%, 90%, 70%, 50%</p> Signup and view all the answers

    What is the final step following the staining process?

    <p>Mounting</p> Signup and view all the answers

    What is the role of eosin in the staining process?

    <p>To stain cytoplasmic structures</p> Signup and view all the answers

    Which of the following best describes a basic stain?

    <p>Stains basic structures and carries a positive charge</p> Signup and view all the answers

    Study Notes

    Light and Electron Microscopy

    • Light Microscope (LM)
      • Uses visible light for illumination
      • Uses glass condensers and lenses for focusing
      • Resolution: 0.2 micrometers, Magnification: up to 4,000x
      • Produces colored images
    • Electron Microscope (EM)
      • Two Types: Transmission Electron Microscope (TEM) and Scanning Electron Microscope (SEM)
      • Uses a beam of electrons for illumination
      • Uses electromagnetic lenses for focusing
      • Resolution: 0.2 nanometers, Magnification: up to 400,000x
      • Produces black and white images

    Staining Techniques

    • Hematoxylin and Eosin (H&E) Stain

      • Most common stain used in histological techniques
      • Hematoxylin: stains nuclei blue (basophilic)
      • Eosin: stains cytoplasm pink (acidophilic)
    • Basic vs Acidic vs Neutral Stains

      • Basic Stains: carry a positive charge (e.g., Hematoxylin, Methylene blue)
      • Acidic Stains: carry a negative charge (e.g., Eosin)
      • Neutral Stains: combination of acidic and basic dyes, stain all tissue components
    • Vital vs Supravital Stains

      • Vital Stains: stain living cells within a living animal (in vivo)
      • Supravital Stains: stain living cells outside the body (in vitro)
    • Trichrome Stains

      • Use three different dyes to differentiate tissue components (e.g., Masson's stain)
      • Differentiate collagen from muscle
      • Masson's Stain: stains collagen green, nuclei blue, and muscle red
    • Metachromatic Stains

      • Stain certain cell components with a color different from the dye used
      • The phenomenon is called metachromasia
      • Example: Toluidine blue staining of mucopolysaccharide granules in mast cells changes to violet

    Histochemical and Immunohistochemical Stains

    • Histochemistry

      • Localizes substances in tissue based on chemical reactions between cell components and stains
      • Reaction produces a permanent colored precipitate
      • Applications: detecting nucleic acids, enzymes, glycogen, and lipids
    • Immunohistochemistry

      • Combines morphology and biochemistry to identify specific antigens
      • Based on antibody-antigen reactions
      • Visualizes the site of reaction microscopically

    Paraffin Technique (Routine Histological Section)

    • Steps:
      • Selection and obtaining tissue
      • Fixation
      • Washing
      • Dehydration
      • Clearing
      • Infiltration and Embedding
      • Microtomy and Sectioning
      • Mounting
      • Deparaffinization
      • Hydration
      • Staining
      • Dehydration
      • Clearing
      • Mounting
      • Cleaning and labeling

    Transmission Electron Microscopy (TEM)

    • Most common stain used for ultrathin sections
    • Heavy metals are used for staining
    • Double contrast method with uranyl acetate (UA) and lead citrate is standard technique
    • Original TEM images are in grayscale

    Types of Microscopes

    • Light Microscopes:
      • Conventional light microscope
      • Polarizing microscope
      • Phase contrast microscope
      • Dark-field microscope
    • Other Microscopes:
      • Fluorescent microscope
      • X-ray microscope

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    Description

    This quiz covers the fundamental concepts of light and electron microscopy, highlighting their differences, magnification, resolution, and image production. It also discusses various staining techniques used in histology, including Hematoxylin and Eosin staining. Test your understanding of microscopy principles and techniques!

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