What is the mechanism of Gram staining?

What is the mechanism of Gram staining?

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 is the role of safranin in Gram staining?

The safranin is also used as a counter-stain in Gram’s staining. In Gram’s staining, the safranin directly stains the bacteria that has been decolorized. With safranin staining, gram-negative bacteria can be easily distinguished from gram-positive bacteria.

What are the steps in a Gram stain and the possible results?

A Gram stain is colored purple. When the stain combines with bacteria in a sample, the bacteria will either stay purple or turn pink or red. If the bacteria stays purple, they are Gram-positive. If the bacteria turns pink or red, they are Gram-negative.

Why methylene blue is used in staining?

It is a cationic dye that stains cells blue because the positively charged dye is attracted to negatively charged particles such as polyphosphates, DNAs, and RNAs. Specimens collected from patients by swabbing are smeared onto microscope slides and the methylene blue solution is dropped on the slide.

Why crystal violet is used in Gram staining?

The gram stain utilizes crystal violet as the primary stain. This basic dye is positively charged and, therefore, adheres to the cell membranes of both gram negative and positive cells. After applying crystal violet and waiting 60 seconds the excess stain is rinsed off with water. Next, a mordant is used.

Why crystal violet is used in gram staining?

What is the most important step in Gram staining?

The thickness of the smear used in the Gram stain will affect the result of the stain. The step that is most crucial in effecting the outcome of the stain is the decolorizing step.

What is Decolorizer in gram staining?

The decolorizer, ethyl alcohol, is the most crtitical step. Ethyl alcohol is a nonpolar solvent, and thus penetrates the cell walls of Gram negative cells more readily and removes the crystal violet-iodine complex.

Why iodine is used in gram staining?

Gram’s iodine is used in Gram staining procedure to differentiate gram positive and gram negative organisms. Gram’s iodine acts as a mordant that causes the crystal violet to penetrate and adhere to the gram –positive organisms.

How do you distinguish between gram-positive and gram-negative bacteria?

Gram positive bacteria have a thick peptidoglycan layer and no outer lipid membrane whilst Gram negative bacteria have a thin peptidoglycan layer and have an outer lipid membrane.

What is the objective of Gram staining?

Introduction: Gram staining is a method commonly used to determine the chemical make up of the cell wall of bacteria. The cell wall can stain either positive or negative, depending on its chemistry. Knowing the chemical make up makes it easier to manipulate the bacteria for various purposes.

Why is gram negative stain red?

Conversely, the the outer membrane of Gram negative bacteria is degraded and the thinner peptidoglycan layer of Gram negative cells is unable to retain the crystal violet-iodine complex and the color is lost. A counterstain, such as the weakly water soluble safranin, is added to the sample, staining it red.

Why mordant is used in gram staining?

After applying crystal violet and waiting 60 seconds the excess stain is rinsed off with water. Next, a mordant is used. The mordant is Gram’s Iodine. This binds to the crystal violet making a large complex that adheres to the cell membrane.