Gram Staining Lab Report

The Gram stain was used to determine if the bacteria was gram positive or negative. A negative test shows a pink color and a positive test is a purple color. When a bacterium is negative it is because it has an outer membrane and a thin layer of peptidoglycan that is much harder to stain. A positive bacterium has a thick layer of peptidoglycan and no outer membrane that can be penetrated by crystal violet.

The next step of the project included preparing a Gram stain to discover the cell shape, arrangement, and if the bacteria is gram positive or

The outer membrane in the cyanobacterium cell (Phormidium uncinatum) is the structure that makes the cell gram negative. On the other hand, the characteristic that makes the cell gram positive is the thick peptidoglycan layer.

This experiment was given to us to utilized previous knowledge learned throughout the semester to identify a gram negative unknown bacterium. We had to first learn the difference between a gram negative and a gram positive organism. We started off with doing gram stains to determine whether it was positive or negative. Based on the gram stains, a gram positive stains purple and a gram negative stains pink. A gram positive stains purple because the cell walls is made of a thick peptidoglycan layer and doesn’t

8. Discuss a possible mechanism of Gram Staining in terms of differences in structure and chemistry between the walls of gram-positive and gram-negative

The purpose of this experiment is to distinguish and indentify an unknown bacterium. There are several tests that can help one eliminate and narrow down the options. The most useful test, and the very first one done, is a gram stain. This test will tell whether the bacterium is gram-positive or gram-negative. After the type of gram stain is identified, the tester has a wide array of differentiating tests at their disposal. Based on the results from these tests, and the numerous others that are available, one can accurately establish the identity of an unknown bacterium.

The chart below shows the biochemical tests of the gram stain below. Test performed Purpose Materials used Observation Results Gram stain test Gram reaction to specimen Crystal violet, iodine, alcohol safranin Pink/purple colored Gram positive cocci/ gram negative rods After the gram stain showed the specimen as Gram positive cocci, and Gram negative rods, more tests are necessary. Test performed Results Enterotube II Gram negative rod: Citrobacter

Gram staining is a technique used to determine if the bacteria is Gram positive or Gram- negative. Gram staining procedure uses crystal violet stain, iodine moderator, alcohol decolorizer and safarin counter stain. In Gram- negative bacteria the primary stain will be washed out with the decolorizer and it will be stained with the counterstain. Whereas in Gram-positive bacteria the primary stain will not leave the cell wall. This difference comes from difference in the structure of the cell wall that retains the stain.

I performed a gram stain on my unknown 7. It appeared to be purple and round shaped. I came to the conclusion that my bacterium is gram-positive, cocci. A gram stain is a differential test. It allows recognizing the difference between two cell walls structures. In a gram-positive cell wall contains a thick peptidoglycan layer covering the plasma membrane (2). The purple staining is retained around the cell

Figure 3. The isolated colonies from figure 2 are viewed under the microscope at the 100x objective after gram staining. In this figure, you can see both gram-negative and gram-positive microorganisms. The gram-negative are the Rhodospirillaceae and they are pink rod-shaped bacteria. The

The gram positive organisms would be purple because the dye remains within the cell; the thick layer of peptidoglycan allows the possession of the purple dye.

From last week’s test, we achieved pure cultural characteristics from the two slants we made. The growth we saw on the agar slant that contained the yellow specimen was a soft, smooth, yellow growth. The growth we saw on the beige specimen was a thin, even, beige growth. Both cultural characteristics were achieved in the appropriate categories. The categories we were looking for contained abundance of growth, pigmentation, optical characteristics, and consistency. Today we will be preparing two bacterial smears from each specimen and Gram staining them. The reason we are conducting this test is to differentiate between two principle groups, gram positive and gram negative and to further know if a pure culture from both organisms was achieved. This is important for classification and differentiation of microorganisms. The Gram stain reaction will help us tell the difference of the chemical composition of bacterial cell walls. The Gram stain procedure uses four different reagents such as crystal violet, gram’s iodine, ethyl alcohol, and safranin. Before the Gram stain is performed we must make two bacterial smears of the two specimens. We placed one loop of distilled water on a clean slide aseptically. He transferred the specimen from the agar slant that contained the yellow growth and placed it on the slide with the water and gently mixed it together in a circular motion approximately the size of a nickel. He let the smear air dry for one minute and gently heat

A gram stain is performed to identify bacteria as gram positive or negative, and to visualize cell arrangement

Escherichia is a genus of aerobic gram-negative rod-shaped bacteria of the family Enterobacteriaceae that form acid and gas on many carbohydrates, such as dextrose and lactose, but not acetone, which include occasional pathogenic forms, including some strains of E. coli which are normally present in the human intestine as well as other forms which typically occur in soil and water (Webster). Escherichia coli is a gram-negative bacilli that rarely varies in shape and size and when stained often resemble safety pins because the ends of some bacilli stain more densely than does the middle; which is a characteristic called bipolar staining which is common in enteric gram-negative bacilli (ASM). Gram negative cells have a thin cell wall layer and will stain red to pink. The staining process is the same as Gram positive, requiring four steps: applying a primary stain, adding a mordant, then rapid decolorization and completing with a counter stain. Applying the alcohol for decolorization dissolves the outer membrane and leaves small holes in the thin peptidoglycan layer through which the crystal violet-iodine diffuse. The gram-negative bacteria is colorless after the decolorization; therefore adding safranin

In all areas of biology, it is easy to see that structure is related to function. This statement holds true in microbiology as well, the study of microorganisms, including bacteria. One characterizing feature of bacteria is the cell wall, which can generally (although not in all situations) be categorized into one of two categories: either Gram positive or Gram negative. Gram positive bacteria’s cell walls are composed of a large peptidoglycan layer (up to 90% of their cell wall). Within this large peptidoglycan layer, one can find techoic acids, which contribute to the maintenance of cell wall structure, and lipotechoic acids, which attach to membrane lipids. Gram positive bacteria that act as pathogens can also potentially release exotoxins, which can have very dangerous effects on humans. Gram negative bacteria, on the other hand, have a very small layer of peptidoglycan in their cell wall, which is surrounded by an outer membrane. Within the outer membrane, one can find the lipopolysaccharide layer, which is one of the most distinguishing factors of Gram-negative bacteria. It is important to note that Gram negative bacteria fail to possess techoic