(* all proper sterilization procedures were practiced throughout this lab*)
NEGATIVE STAIN
Materials used for the Negative stain:
1. 2 clean microscope slides that will be used for the staining process.
2. 2 clean microscope slides that will be used to spread the Nigrosin.
3. Nigrosin stain.
4. Staining rack over the sink.
5. Disinfectant solution.
The steps that were taken for both the environmental sample slide and for the unknown sample:
1. We placed a small drop of Nigrosin near one end of the slide.
2. With a loop, we transferred a small amount of bacteria from an agar plate into the Nigrosin drop and mixed the bacteria and Nigrosin together.
3. We touched (and slid) the short edge of another clean slide to the slide with the contaminated Nigrosin at a 45 degree angle.
4. After the contaminated Nigrosin drop was spread along the edge of the spreader slide, we quickly pulled the slide to spread out the drop.
5. We allowed the smear to air dry completely.
6. We repeat this process for the second slide
CAPSULE STAIN
Materials used for the capsule stain:
1. negative stained slides
2. safranin or crystal violet
3. distilled water
This stain is used to detect whether or not either of our environmental bacteria or unknown bacteria have capsules.
1. We took our prepared negative stains and, after allowing them to completely dry, stained our environmental stain with safranin and our unknown with crystal violet.
2. After allowing the stain to set in for about a minute each, we gently washed off the excess stain with distilled water.
3. Blot the water from the slide with bibulous paper.
4. We examined the smears under the microscope using the oil immersion lens.
5.We washed the slides in disinfectant solution
Above Results: Our unknown sample = bacilli shaped, no capsule
Above Results:
Environmental sample= Rod-shaped, no capsule
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