Microscopic Evaluation: Common Stains and Techniques
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Questions and Answers

Which fluorochrome stain is used to screen for acid-fast bacilli and produces a yellow or orange fluorescence under a fluorescent microscope?

  • Auramine-rhodamine (correct)
  • Methylene Blue
  • Acridine Orange
  • Calcofluor White
  • Which stain is used to visualize the capsule surrounding certain yeasts, such as Cryptococcus species, by creating a dark background and a clear capsule around the yeast?

  • India Ink (correct)
  • Endospore Stain
  • Acridine Orange
  • Lactophenol Cotton Blue
  • Which stain is used to stain the cell walls of medically important fungi grown in slide culture?

  • Lactophenol Cotton Blue (correct)
  • Auramine-rhodamine
  • Methylene Blue
  • Calcofluor White
  • Which stain is used to stain Corynebacterium diphtheriae for observation of metachromatic granules?

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

    Which stain is used to visualize endospores, where the endospores appear green within pink- or red-appearing bacterial cells?

    <p>Endospore Stain</p> Signup and view all the answers

    Which stain is a fluorochrome that binds to chitin in fungal cell walls, producing a bright apple-green or blue-white fluorescence?

    <p>Calcofluor White</p> Signup and view all the answers

    Which of the following is the correct procedure for preparing a thin, monolayer smear?

    <p>Roll a swab back and forth across the slide to create the smear</p> Signup and view all the answers

    Which of the following stains is used to specifically identify organisms?

    <p>Probe-mediated stains</p> Signup and view all the answers

    What is the purpose of the Gram stain in a clinical microbiology laboratory?

    <p>To determine the cell wall structure and Gram-staining characteristics of organisms</p> Signup and view all the answers

    What is the purpose of the acid-fast stain?

    <p>To identify organisms that are resistant to decolorization by acidic alcohol</p> Signup and view all the answers

    Which of the following stains is commonly used in diagnostic microbiology laboratories?

    <p>All of the above</p> Signup and view all the answers

    Which of the following is NOT a step in the Gram staining procedure?

    <p>Heating the primary stain</p> Signup and view all the answers

    What is the primary purpose of the Gram staining technique?

    <p>To differentiate between Gram-positive and Gram-negative bacteria</p> Signup and view all the answers

    Which of the following statements about Gram-negative bacteria staining is correct?

    <p>Pseudomonads, anaerobic bacilli, and thin-walled bacteria stain poorly with safranin</p> Signup and view all the answers

    Which staining method is specifically designed for bacteria with high lipid and wax content in their cell walls?

    <p>Acid-fast staining</p> Signup and view all the answers

    What is the difference between the Ziehl-Neelsen and Kinyoun acid-fast staining methods?

    <p>The Ziehl-Neelsen method uses heat, while the Kinyoun method is a cold or detergent method</p> Signup and view all the answers

    Which of the following organisms can be identified using the acid-fast staining technique?

    <p>Mycobacteria, yeast, and coccidian parasites</p> Signup and view all the answers

    Study Notes

    Staining Procedures

    • Decolorization: use 3.0% acid-alcohol (faint pink color remains)
    • Counterstaining: Loeffler's methylene blue
    • Washing: with water and deionized water

    Common Stains Used for Microscopic Evaluation

    • Auramine-rhodamine: fluorescent stain for acid-fast bacilli, appears yellow or orange under fluorescent microscope
    • Acridine Orange: fluorochrome dye for gram-positive and gram-negative bacteria, appears bright orange under fluorescent microscope
    • Calcofluor White: fluorochrome stain for fungal cell walls, appears bright apple-green or blue-white fluorescence
    • Methylene Blue: for staining Corynebacterium diphtheriae to observe metachromatic granules
    • Lactophenol Cotton Blue: for staining cell walls of medically important fungi grown in slide culture
    • India Ink: negative stain for visualizing capsules surrounding certain yeasts, such as Cryptococcus spp.
    • Endospore Stain: malachite green and safranin for visualizing endospores in bacterial cells

    Gram Staining

    • Results:
    • Gram-positive bacteria: primary stain crystal violet appears dark blue to blue-black
    • Gram-negative bacteria: counterstain safranin appears bright red
    • Enterics have strong avidity and stain bright red
    • Pseudomonads stain moderately well
    • Anaerobic bacilli and other thin-walled bacteria stain weakly

    Acid-Fast Staining

    • Used for bacteria with high lipid and wax content in their cell walls
    • Two methods: Ziehl-Neelsen stain (hot method) and Kinyoun stain (cold method)
    • Used to identify Mycobacteria, yeast, Saccharomyces, and coccidian parasites such as Isospora belli, Cryptosporidium, and other coccidia-like bodies.

    Categories of Stains

    • Simple stains: forms and shapes
    • Differential stains: specific components of elements present
    • Probe-mediated stains: diagnostic antibody or DNA probe-mediated stains for identification of organisms

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    Description

    This quiz covers the staining techniques used in microscopic evaluation, including decolorization, counterstaining, and common stains like Auramine-rhodamine and Acridine Orange. Test your knowledge on different staining methods for bacteria.

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