Why would gram negative bacteria appear purple after gram stain?

Answer 1

The simple staining procedure makes to visualize bacteria clearly, but it does not distinguish between organisms of similar morphology. The Gram staining method is named after the Danish bacteriologist (1882) Hans Christian Gram, is one of the most important staining techniques in microbiology. It is almost always the first test performed for the identification of bacteria. 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. These microorganisms that are stained by the Gram's method are commonly classified as Gram-positive or Gram non-negative. Others that are not stained by crystal violet are referred to as Gram negative, and appear red. In this method the fixed bacterial smear is subjected to the following staining reagents in the order of sequence listed below Primary stain Crystal violet is used first and stains all cell purple = Mordant = Grams iodine serves as a mordant, a substance that increases the cell affinity for a stain. It does this by binding to the primary stain thus forming an insoluble complex. The resultant crystal violet iodine complex serves to identify the color of the stain. At this point all cells will appear purple black. = Decolorizing agent = Gram staining is based on the ability of bacteria cell wall to retaining the crystal violet dye during solvent treatment. The cell walls for Gram-positive microorganisms have a higher peptidoglycan and lower lipid content than gram-negative bacteria In Gram-negative cells, subsequent treatment with a decolorizer, (95% ethyl alcohol) dissolves the lipid layer from the gram-negative cells. The removal of the lipid layer enhances the leaching of the primary stain from the cells into the surrounding solvent. In contrast, the solvent dehydrates the thicker Gram-positive cell walls, closing the pores as the cell wall shrinks during dehydration. As a result, the diffusion of the violet-iodine complex is blocked, and the bacteria remain stained. Counter stain A counterstain of basic fuchsin or saffranin is applied to the smear to give decolorized gram-negative bacteria a pink color. Since only gram-negative cells undergo decolorization they may now absorb the counter stain. Gram-positive cell retain the purple color of the primary stain. Basic fuchsin stains many Gram-negative bacteria more intensely than does safranin, making them easier to see. Some bacteria which are poorly stained by safranin, such as Haemophilus spp., Legionella spp., and some anaerobic bacteria, are readily stained by basic fuchsin, but not safranin. The polychromatic nature of the gram stain enables determination of the size and shape of both Gram-negative and Gram-positive bacteria. If desired, the slides can be permanently mounted and preserved for record keeping.

Answer 2

When we stain bacteria we use a substance called Gram's stain. The bacteria take up the red or the purple in the stain depending on their type. Gram positive bacteria picks up the purple portion in the gram stain and the gram negative pick up the red color of the stain. As an example, Staphylococcus and Streptococcus are gram positive bacteria while E.which is the most common bacteria in urinary tract infections are gram negative. It should be noted that many food borne illnesses through out the country have been infected with E. coli, which is also known as Escherichia coli the genus and species of the bacteria.

Answer 3

They are not naturally purple.

The gram stain used to test for them is a purple dye that binds to the cell wall of gram positive bacteria but not to the cell wall of gram negative bacteria. Once this stain is applied they become purple.

Answer 4

CAN