Smear Preparation
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Questions and Answers

What is the purpose of adding a loopful of water to the slide in smear preparation?

  • To stain the bacteria
  • To fix the bacteria to the slide
  • To create a suspension of bacteria (correct)
  • To kill the bacteria

After air drying a smear, what is the next step in smear preparation?

  • Staining
  • Cover slipping
  • Fixing (correct)
  • Washing

Why is heat fixing performed on a smear?

  • To kill all the bacteria
  • To adhere the bacteria to the slide (correct)
  • To make the bacteria bigger
  • To make the bacteria move

Before heat fixing, how long should a smear air dry?

<p>Less than 10 minutes (B)</p> Signup and view all the answers

What is the primary purpose of staining microorganisms?

<p>Enhancing the contrast for better visualization (A)</p> Signup and view all the answers

What is a differential stain used to do?

<p>Identify different types of cells (D)</p> Signup and view all the answers

What is the first step in Gram staining after preparing a smear?

<p>Adding crystal violet (B)</p> Signup and view all the answers

What color are Gram-positive bacteria after staining with crystal violet?

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

Which reagent acts as a mordant in the Gram staining procedure?

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

What is the purpose of alcohol in Gram staining?

<p>To decolorize Gram-negative bacteria (A)</p> Signup and view all the answers

What color are gram-negative bacteria after the addition of safranin?

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

Safranin functions as what in the Gram stain procedure?

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

What color do Gram-positive bacteria appear after Gram staining?

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

What is a characteristic of acid-fast positive cells?

<p>They retain a basic stain in the presence of acid-alcohol (B)</p> Signup and view all the answers

What happens to non-acid-fast cells during acid-fast staining?

<p>They lose the primary stain and are counterstained (B)</p> Signup and view all the answers

What cell wall component makes cells difficult to stain with Gram stain?

<p>Mycolic acid (D)</p> Signup and view all the answers

What makes bacteria impermeable to staining procedures, including the Gram stain?

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

Why are some bacteria called 'acid-fast'?

<p>They resist decolorization with acid alcohol (D)</p> Signup and view all the answers

Which of the following is a mordant used in the Ziehl-Neelsen method?

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

In the Ziehl-Neelsen method, what is used as the counterstain?

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

What color will acid-fast cells be after staining with the Ziehl-Neelsen method?

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

In the Kinyoun method, what replaces heat?

<p>Higher concentration of phenol (D)</p> Signup and view all the answers

In the Kinyoun method, what is used as the counterstain?

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

What color will non-acid-fast cells be after staining with the Kinyoun method?

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

What is the primary stain used in both the Ziehl-Neelsen and Kinyoun methods?

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

What decolorizer is used in the acid-fast stain?

<p>Acid-alcohol (A)</p> Signup and view all the answers

What is the purpose of using acid-alcohol in acid-fast staining?

<p>To decolorize non-acid-fast cells (B)</p> Signup and view all the answers

After completing the Ziehl-Neelsen staining process, what color will acid-fast cells appear?

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

What is the determining factor of virulence for the bacterium M. Tuberculosis?

<p>Mycolic acids (A)</p> Signup and view all the answers

Mycobacterium tuberculosis is the causative agent of which disease?

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

Which cell wall component makes bacteria acid-fast?

<p>Mycolic acid (D)</p> Signup and view all the answers

Which of the following is a non-pathogenic Mycobacterium species?

<p><em>Mycobacterium smegmatis</em> (C)</p> Signup and view all the answers

What is the primary stain in the Gram stain?

<p>Crystal Violet (A)</p> Signup and view all the answers

In the Gram stain, what is the function of Gram's iodine?

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

Which step differentiates Gram-positive from Gram-negative bacteria?

<p>Decolorizing with alcohol (A)</p> Signup and view all the answers

What is the correct order of reagents in the Gram stain procedure?

<p>Crystal violet, iodine, alcohol, safranin (C)</p> Signup and view all the answers

Which of the following is a differential stain?

<p>Gram stain (A)</p> Signup and view all the answers

What is the purpose of heat in the Ziehl-Neelsen method?

<p>To drive the stain into the cell (A)</p> Signup and view all the answers

What is the counterstain used in Ziehl-Neelsen staining?

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

What is the primary stain in the acid-fast staining?

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

What is the function of acid-alcohol in acid-fast stain?

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

What is added to the slide before adding the inoculum in a smear preparation?

<p>A loopful of water (C)</p> Signup and view all the answers

In the Ziehl-Neelsen method, what role does heat play?

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

What component of the bacterial cell wall makes it difficult to stain with Gram stain?

<p>Mycolic acid (D)</p> Signup and view all the answers

What decolorizer is used in acid-fast staining?

<p>Acid-alcohol (B)</p> Signup and view all the answers

What color will non-acid-fast cells appear at the end of the Ziehl-Neelsen staining process?

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

Flashcards

Smear preparation

A procedure to create a bacterial suspension on a slide for staining.

Loopful of water purpose

Adding a small amount of water to the slide to create a suspension.

Inoculum in smear prep

Adding bacteria to the drop of water on the slide to create a suspension.

Air drying a smear

Important to dry the slide before applying heat.

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Heat fixing a smear

The process of gently heating the air-dried smear to kill bacteria and fix them to the slide.

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Differential stain

A staining method that differentiates bacteria based on their cell wall structure.

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Acid-fast cells

Cells that retain a basic stain despite acid-alcohol washes.

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Non-acid-fast cells

Cells that lose the primary stain when rinsed with acid-alcohol and take up the counterstain.

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Acid-fast stain

A differential stain used to identify bacteria with waxy cell walls.

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Mycolic acid

A waxy substance in acid-fast bacteria cell walls that makes them impermeable to many stains.

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Ziehl-Neelsen method

The primary stain used in this method is carbol fuchsin, apply heat as a mordant, uses acid-alcohol to decolorize, and methylene blue as counterstain. Acid-fast cells appear red, non-acid-fast cells appear blue.

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Kinyoun method

This method uses a higher concentration of phenol in carbol fuchsin, no heat as mordant, uses acid-alcohol to decolorize, and brilliant green as counterstain. Acid-fast cells appear red, non-acid-fast appear green.

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Acid-alcohol

A solvent used to remove the primary stain from non-acid-fast cells.

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Counterstain

A stain used to colorize decolorized cells, providing contrast.

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Purpose of heat-fixing

To kill and attach the bacteria to the slide

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Heat as a mordant

The mordant in the ZN method used to increase stain penetration.

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Methylene blue in ZN

Dye used to stain non-acid-fast cells in the ZN method.

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Mycobacterium tuberculosis

Pathogenic species; causes tuberculosis.

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Tuberculosis (TB)

A bacterial infection and a leading cause of death worldwide.

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Mycolic Acids in MTB

Thought to significantly determine virulence in MTB.

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Mycobacterium avium

Genus include Mycobacterium avium which causes a TB-like disease.

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Carbol Fuchsin

This is the primary stain used during the acid-fast stain (ZN method)

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Steaming with Carbol Fuchsin

Heating gently until slight steaming; maintain for 5 min, adding more to prevent drying.

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Acid fast stain color

What color do acid-fast cells stain in the Ziehl-Neelsen method at the end of the staining?

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Non Acid fast stain color

What color do non-acid-fast cells stain in the Ziehl-Neelsen method at the end of the staining?

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Study Notes

Smear Preparation

  • Add a loopful of water to a slide to create a bacterial suspension.
  • Add the inoculum to the water drop, creating a suspension on the slide.
  • Let the smear air dry.

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Description

A quick guide to preparing a bacterial smear on a slide. This involves creating a bacterial suspension with water, adding inoculum, and allowing the smear to air dry. Ideal for microbiology students.

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