In a clinical setting signs and symptoms can lead a care taker down a treatment path, but if a bacterial infection is suspected, a definitive answer from the “lab” is needed before prescribing medications. In this instance a yellow sputum was produced by the patient, which will be taken to the lab an analyzed under a microscope. There are three suspect bacteria, which are Bacillus, Mycoplasma, and Escherichia. A discussion of staining procedures, as well as anatomical differences will be discussed for each specimen.
Bacillus are Prokaryotic cells, that are rod shaped bacteria and can from long chains. Bacillus are also spore forming. When gram staining a Bacillus sample, they are found to be gram positive. However, there are some Bacillus that are gram negative.
Escherichia are similar to Bacillus, in that they are rod shaped, but they are non spore forming. The significance in this is that Bacillus spores can infect up to hundreds if not thousands of years after they were produced, as they are harbored in soil. Most Escherichia are motile by flagella and are gram negative
Mycoplasma are the smallest known bacteria. They are so small that they will pass through filters that most bacteria could not pass through. Mycoplasma does not have a cell wall, which allows for it to take many different shapes. The lipids in the plasma membrane in the cell creates sterols, which protect it from rupture. The peptioglyan proteins are a target for antimicrobial activity. It disrupts the final linking of the peptidoglycan rows, which leads to it's destruction due to rupture (Tortora, 2010, pg 85). Mycoplasma are gram positive when stained.
To prepare the sample for staining and stain the sample the following steps would be taken:
Take a small sample of sputum and smear it on a slide. This is then tempered by heat on to the slide, which will prevent washing it off with the stains.
Crystal violet, a primary stain, is placed