1) a non-acid fast control (M. Luteus)
2) soil microbe
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| M. Luteus (non-acid fast control) |
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| Soil Microbe |
During this acid stain procedure, we used an aseptic technique and smeared a sample of our soil microbe and the control culture onto a slide, along with a loopful of water. We mixed the bacteria with the water and allowed it to dry. Then, we put our slide through the bunsen burner and placed the slides over a beaker with steaming water. The slides were then flooded with carbol-fushin and cooled/rinsed with water. The slides were depolarized with acid-alcohol until the color stopped coming off. Lastly, the slide was flooded with methylene.
Below is a picture of the control slide & soil microbe over steaming water:
We then observed our acid fast stain slides under the microscope at 100X magnification. Sidenote: (Oil immersion was added to the slide).
Below is a picture of the control and our soil microbe under the microscope:
The soil microbe had a blue tint with many blue rods (bacilli) distributed across the slide.
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| Soil Microbe |
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| M. Luteus |
It is important to include some background information about an acid fast stain and what it actually is to further understand and interpret our results. An acid fast stain is a stain that is used to identify organisms, such as members of the genus, Mycobacterium. Acid fast organisms usually have waxy cell walls that are impermeable and contain many lipids and fatty acids. Usually to get through this cell wall, heat is involved.
It was found that our soil microbe, was non-acid fast, which means that it does not have a waxy cell wall. It was also gram (+) in terms that a peptidoglycan wall is present.
Dichotomous Key
Below, I have used a dichotomous key in order to begin identifying our microbe…Below is the steps that I followed in the dichotomous key:
1. Gram Positive
2. Morphology, Rod Shaped (Bacilli)
3. Non-acid fast organism
4. …The next question asks if the organism is catalase (+) or catalase (-)…Unfortunately, at this time in the experiment we do not know the answer to this question yet so we will have to look forward to next week's lab in order to further identify our mysterious microbe!
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