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by: Jenna Hill

Practice Random

Jenna Hill


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This 2 page Class Notes was uploaded by Jenna Hill on Monday September 26, 2016. The Class Notes belongs to Random at Montana State University - Bozeman taught by in Fall 2016. Since its upload, it has received 4 views.


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Date Created: 09/26/16
Jenna Marie Hill Microbe Lab 1:40­2:30 1. Paramecium Aurelia, also known as protozoa, are eukaryotic, heterotrophic  microorganisms that are considered to be animal­like. They are found in water samples,  such as pond water, which requires a wet mount slide. To make a wet mount slide, you  will first need gather the following materials: the sample, a clean glass slide, a cover slip,  and a sterile eye dropper (I didn’t know what else to call it). The first step in making a  wet mount slide is get the sterile eye dropper ready, (be sure not to touch the tip), and  draw a small amount of your sample. Next, get a clean glass slide and place a small  amount of the sample on the slide. Now, take a cover slip, place it on its edge, slanted  over the sample, and let it fall. Using this technique helps decrease the amount of bubbles on your slide. When you place your slide under the microscope, start with the lowest  power lens, 10x. This gives you the ability to find a nice spot where there could be some  microorganisms. When you find a good spot, move up to the next lens, 40x. Since  protozoa are bigger, they can be seen with the 40x lens. Protozoa look like clear jelly  beans with tiny, sometimes clear dots inside them. These tiny dots inside them are  organelles that help the protozoa function. You can often witness them look for, and  consume other microorganisms in their environment. Staphylococcus aureus is a gram positive bacterium that is commonly found in the nose,  in the respiratory tract, and on the skin. It is the common cause of skin infections,  respiratory infections, and food poisoning. Since Staphylococcus aureus is a bacterium,  you will need to make a gram stain of you sample in order to see anything, since bacteria  are clear and microscopic. To make a gram stain, you will need to gather the following  materials: a clean glass slide, an inoculation loop, your sample, an autoclave, and the  stains needed. The first step to making a gram stain is to get the glass slide, and place a  small amount of water on it. Next, sterilize the inoculating loop in the autoclave and let it  cool so that you don’t kill you sample. Gather a small colony from you sample, or dip  your loop in the liquid sample, and swirl it around in the water that you placed on the  glass slide earlier. Then lay the slide on top of the autoclave to let the water evaporate,  leaving just the sample. After the water has completely evaporated, it is time to start  staining. First, soak the slide in crystal violet for 1 minute. Crystal violet stains  everything on you slide purple/blue, which allows you to see the bacteria. After rinsing  the crystal violet off with distilled water, soak your slide with iodine for 1 minute, then  rinse. The iodine forms a complex with the crystal violet, which makes it extremely  difficult to remove from the gram positive organisms. Next, flood the slide with alcohol  for just a few seconds, then rinse. The alcohol removes the stain from the gram positive  organisms, which have a thinner peptidoglycan layer than the gram positive. The last step is to flood the slide with safranin for 30 seconds, then rinse. The safranin stains the now  colorless gram negative organisms, so that they can now be seen under the microscope.  Dry slide off with bibulous paper. Now you can place you slide under the microscope. I  start off with the 40x lens so that I can find a nice spot before placing the oil on the slide  for the 100x lens. After you have found a nice spot, place some oil on your slide and put  the 100x lens in place. Since Staphylococcus aureus is a gram positive organism,  meaning it has a thick layer of peptidoglycan, it will show up as deep purple, grape­like  clusters.  Cladosporium orchidis is a Cladosporium fungus found on orchids. It appears as olive­ green spots on plants that enters through local damage from things such as frost. The  spores are dispersed by wind and can cause infections of the skin, toenail, sinus, and  lungs. Since Cladosporium orchidis is a fungus, you will need to make a tape mount slide in order to view your sample under the microscope. To make a tape mount, you will need clear tape, and a glass slide. Take a piece of tape, gently touch the outer edge of you  sample (to gather a more spread out sample), and place the tape, sticky side down on the  slide. Place your slide under the microscope, and start out at 10x to find a nice spot on  your slide. When you have found a good spot, move up to 40x. The fungus is large  enough that you do not need a higher power lens. Cladosporium orchidis looks like a  tiny, olive­green, dead tree, branching outwards. Arthrospira platensis, also known as Spirulina, is a gram negative bacterium that is a  species of blue­green cyanobacteria. It is nontoxic, and is actually very nutritious. People  in tropical areas are known to eat it, and it has many health benefits, such as lower blood  pressure, and reduced cholesterol. It is also rich in vitamins, minerals, beta­carotene,  essential fatty acids, and antioxidants. Since it is a bacterium, you will need to gram stain  a sample in order to view it under the microscope. (Since I already described, in detail,  the process of gram staining when I talked about Staphylococcus aureus, I will not go  into detail about it here). When your sample is ready, place it under the microscope, find  a nice spot using the 40x lens. Then apply the oil and move into the 100x lens. Spirulina  are multicellular, and the individual cells are rectangular in shape, and connected in  chains about 5 cells long. They are connected so that the cells are wider than they are  long. They are usually green in color, and sometimes have a spiral shape, like a cork  screw. 2. You can tell your microscope needs to be cleaned when you are looking at a sample, and  there is a big, blurry blob in your vision and it doesn’t move when you use the travel  knobs. The best way to clean it is to take lens paper, wipe the ocular lens first, then move  to the lowest power lens, and work your way up to the highest power. Leave the 100x for  last since it is covered in oil and you don’t want to spread the oil to the other lens.  3. I think my favorite organism I have seen is the spirilla shaped bacterium. I really like the  way that it moves through its environment. It just swirls around, doing its little bacteria  thing. I didn’t expect to see it when I was zooming around looking at bacteria, but when I found it, it was pretty exciting. It was among a bunch of cocci (of a variety of cell  arrangement) and a few diplo/streptobacilli.


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