Acid-Fast Staining Procedure Quiz
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Questions and Answers

What is the primary purpose of acid-fast staining?

  • To differentiate between Gram-positive and Gram-negative bacteria
  • To enhance bacterial growth
  • To identify acid-fast microorganisms (correct)
  • To identify spore-forming bacteria

Which reagent is NOT used in acid-fast staining?

  • Alkaline methylene blue
  • Methanol (correct)
  • Carbolfuschin
  • Acid alcohol

What color do acid-fast bacilli appear after the staining process?

  • Green
  • Blue
  • Bright red (correct)
  • Brown

During decolorization, what indicates that the slide has been adequately treated?

<p>The slide remains slightly pink (D)</p> Signup and view all the answers

Why is it essential to prevent boiling during the staining process?

<p>It prevents the specimen from being destroyed (A)</p> Signup and view all the answers

How long should the carbolfuschin be heated on the slide during staining?

<p>3-5 minutes (A)</p> Signup and view all the answers

What is the correct order of steps after rinsing with water following decolorization?

<p>Rinse, counterstain, blot dry (D)</p> Signup and view all the answers

What type of microscope objective should be used to examine the slide after staining?

<p>Oil immersion objective (C)</p> Signup and view all the answers

Which of the following bacteria can also be identified through acid-fast staining besides Mycobacteria?

<p>Nocardia (B)</p> Signup and view all the answers

What is the consequence of allowing the slide to dry out during the staining process?

<p>The staining will be ineffective (D)</p> Signup and view all the answers

Flashcards

Acid-Fast Staining

A staining technique used to identify bacteria with a waxy cell wall.

Mycobacteria

Mycobacteria species are a type of bacteria that possess a waxy cell wall, making them difficult to stain with traditional methods.

Heat-Fixing

The process of heating a smear during staining to enhance the penetration of dyes.

Ziehl's Carbolfuschin

A primary stain used in acid-fast staining consisting of carbolfuchsin, which gives the acid-fast bacteria a red color.

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Acid-Alcohol

A decolorizing solution consisting of acid and alcohol. It removes the primary stain from non-acid-fast bacteria but not from those with a waxy cell wall.

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Methylene Blue

A secondary stain used in acid-fast staining that is used as a counterstain to color the background and non-acid-fast bacteria.

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Acid-Fastness

The ability of a dye to be retained in the presence of acid-alcohol

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Acid-Fast Bacilli

Acid-fast bacteria appear red under the microscope.

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Nocardia

A bacteria genus that includes Nocardia, which is known to be

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Oil Immersion

The process of examining a slide under a microscope using immersion oil to improve resolution.

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Study Notes

Acid-Fast Staining Procedure

  • Acid-fast staining identifies acid-fast microorganisms, like Mycobacteria species, and some other bacteria (e.g., Nocardia).
  • Also known as Ziehl-Neelsen staining.
  • Uses carbolfuschin, acid alcohol, and methylene blue.
  • Acid-fast bacilli appear bright red after staining.

Procedure Steps

  • Smear preparation: Mix microorganisms and air dry, then heat-fix the smear.
  • Carbol-fuschin application: Place slide on a heated plate (covered with paper towel) and saturate with carbolfuschin. Heat for 3-5 minutes, avoiding drying or boiling. Alternatively, use a water bath with staining rack.
  • Water rinse: Rinse the slide with water for 30 seconds.
  • Decolorization: Carefully add acid-alcohol dropwise until slight pink remains (10-30 seconds).
  • Second water rinse: Rinse with water for 5 seconds.
  • Counterstain: Stain with alkaline methylene blue for 2 minutes.
  • Final water rinse: Rinse with water for 30 seconds.
  • Drying: Blot the slide dry with bibulous paper.
  • Observation: Examine under oil immersion.
  • Result interpretation: Acid-fast organisms are red; other organisms are blue or brown.

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Description

Test your knowledge on the acid-fast staining procedure, a vital technique for identifying acid-fast microorganisms such as Mycobacteria. This quiz covers the key steps, reagents used, and the significance of the staining results. Enhance your understanding of microbiology techniques with this focused assessment.

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