What are the steps of the Gram stain procedure?
The performance of the Gram Stain on any sample requires 4 basic steps that include applying a primary stain (crystal violet) to a heat-fixed smear, followed by the addition of a mordant (Gram’s Iodine), rapid decolorization with alcohol, acetone, or a mixture of alcohol and acetone and lastly, counterstaining with …
What is Gram staining and explain its procedure?
The Gram stain involves staining bacteria, fixing the color with a mordant, decolorizing the cells, and applying a counterstain. The primary stain (crystal violet) binds to peptidoglycan, coloring cells purple.
How do you prepare a Gram stain slide?
There are six basic steps:
- Apply a smear of bacteria on to a slide.
- Add about 5 drops of Hucker’s Crystal Violet to the culture.
- Add about 5 drops of iodine solution to the culture.
- Tilt slide and decolorize with solvent (acetone-alcohol solution) until purple color stops running.
- Add about 5 drops of Safranine O.
Why do we perform gram staining?
Gram staining differentiates bacteria by the chemical and physical properties of their cell walls. Gram-positive cells have a thick layer of peptidoglycan in the cell wall that retains the primary stain, crystal violet.
What is the most critical step in the gram staining procedure?
The critical step of the Gram staining procedure is the decolorization step. Hold the slide in a tilted downward position and allow the decolorizer to flow over the smear.
How do you prepare a slide for staining?
To prepare the slide:
- Place a drop of fluid in the center of the slide.
- Position sample on liquid, using tweezers.
- At an angle, place one side of the cover slip against the slide making contact with outer edge of the liquid drop.
- Lower the cover slowly, avoiding air bubbles.
- Remove excess water with the paper towel.
What is the principle of Gram stain?
The basic principle of gram staining involves the ability of the bacterial cell wall to retain the crystal violet dye during solvent treatment. Gram-positive microorganisms have higher peptidoglycan content, whereas gram-negative organisms have higher lipid content.
What are the four stages of gram staining?
The Gram staining process includes four basic steps, including:
- Applying a primary stain (crystal violet).
- Adding a mordant (Gram’s iodine).
- Rapid decolorization with ethanol, acetone or a mixture of both.
- Counterstaining with safranin.
What are principles of gram staining?
What is the principle of gram staining?
What is the purpose of gram staining?
A Gram stain is a laboratory test that checks for bacteria at the site of a suspected infection or in certain bodily fluids. A medical laboratory scientist processes the Gram stain, which gives relatively quick results, so healthcare providers can know if bacteria are present, and, if so, the general type(s).
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