Activity 2 Gram Stain
Activity 2 Gram Stain
Instruction:
Guide Questions:
4. A student completed the gram stain technique but neglected to use the
5. What are the factors that will affect the staining affinity of bacteria?
reaction? Discuss.
7. Why isn’t gram stain used on acid-fast bacteria? If you did gram stain
acid-fast bacteria, what would their gram reaction be? What is the gram
ANSWERS:
1. Gram's stain remains one of the most useful tools we have for the accurate
If everyone was using different methods so you couldn't be 100 percent sure
that the outcomes of each were the same. The colors, highlights, times and all
are standardized so that the same results are obtained for everybody who
does it.
If some of the measures were out of place the staining would not work
properly.
stain would not be able to interact with gram-positive bacteria (or less
likely to).
coat a structure (such as a flagellum) to make it thicker and easier to see after
3. For me the procedure is not simple because there are several factors that
may affect the result of gram staining like for example: If the smear is too
during heat fixing, the cell walls will rupture. Concentration and freshness of
reagents may affect the quality of the stain. It is not a simple stain. A simple
stain is when you use just one dye; just one stain to colorize the cells.
process, the gram-positive organisms will not show up violet, they will instead
5. The factors that will affect the staining affinity of bacteria are the following;
Concentration of the dye - The greater the concentration of the dye, the
temperature increases the rate at which the dye diffuses throughout the tissue
sample. It can also alter tissue components so that they are more receptive to
Therefore, the pH of the staining solution can have a direct impact on the
ability of a dye to bind with its intended tissue element. Tissue fixation -
Fixation alters and reorganizes certain molecular structures within the tissue
sample such that there is increased permeability for staining. Unfixed tissue
elements have limited binding sites for dyes. Mordants - Mordants are
chemicals that may be needed to bind dyes to certain tissue elements. They
6. Gram staining differentiates the bacteria from their cell walls by the
chemical and physical properties. Gram-positive cells in the cell wall have a
thick coating of peptidoglycan, which contains the primary dye, crystal violet.
crystal violet to wash out when ethanol is applied. The counterstain, usually
safranin, or fuchsine, stains them pink or red. Upon addition of crystal violet,
Lugol's iodine solution is often applied to reinforce the stain 's bonds with the
cell membrane. Gram staining is almost always the first step in recognizing a
tool in both clinical and research settings, not all bacteria can be definitively
indeterminate groups.
waxy lipid within the structure of their cell walls. Due to the presence of this
lipid, water-based stains, such as the gram stain, so not work well on acid-fast
organisms. If gram stain did in acid fast bacteria, nothing will happen. the