What is stain reagent?

What is stain reagent?

In the staining technique, cells on a microscope slide are heat-fixed and stained with a basic dye, Crystal Violet, which stains all bacterial cells blue. Iodide solution is then added that allows the iodine to enter the cells and form a water-insoluble complex with the Crystal Violet dye.

What are the reagents used in Gram staining and their functions?

The first reagent is called the primary stain. Its function is to impart its color to all cells. The second stain is a mordant used to in- tensify the color of the primary stain. In order to es- tablish a color contrast, the third reagent used is the decolorizing agent.

How do you make a Gram stain reagent?

Gram Iodine Solution: Dissolve 1 g of iodine, 2 g of potassium iodide, and 3 g of sodium bicarbonate in 300 ml of water. Gram Decolorizer Solution: Mix equal volumes of 95 % ethanol and acetone. Gram Safranin Solution: Dissolve 2.5 g of safranin O in 100 ml of 95 % ethanol to make a stock solution.

What can be used instead of safranin?

basic fuchsin
Interpretation of Results If basic fuchsin is used instead of safranin, gram- positive cells appear bright purple to purplish-black, while gram-negative cells appear bright pink to fuchsia.

What infections are caused by Gram positive bacteria?

Streptococcus pyogenes is a gram-positive group A cocci that can cause pyogenic infections (pharyngitis, cellulitis, impetigo, erysipelas), toxigenic infections (scarlet fever, necrotizing fasciitis), and immunologic infections (glomerulonephritis and rheumatic fever).

What are the reagents used in the Endospore stain?

Endospore staining techniques are classified based on the types of reagents used; Schaeffer Fulton Stain- used Malachite Green dye and safranin. Dorner method of endospore staining –uses Carbolfuchsin stain, acid alcohol, and Nigrosin solution)

What reagents could be used in Gram stain?

The Gram stain reagents, listed as they are used in the staining procedure, are crystal violet, iodine, ethanol/acetone and safranin. The initial reaction of the staining procedure is with the addition of the primary stain, crystal violet, to the bacterial smear.

What is the most important reagent in Gram staining?

Furthermore, what is the most important reagent in Gram staining? The primary stain of the Gram’s method is crystal violet. Crystal violet is sometimes substituted with methylene blue, which is equally effective. The microorganisms that retain the crystal violet-iodine complex appear purple brown under microscopic examination.

What is the Order of reagents in the Gram stain?

Reagents: Crystal violet (primary stain) Iodine solution/Gram’s Iodine (mordant that fixes crystal violet to cell wall) Decolorizer (e.g. ethanol) Safranin (secondary stain) Water (preferably in a squirt bottle)

What results do you get with a Gram stain?

Gram Positive : Dark purple