Objective and Purpose:

The objectives and purposes of the lab were to practice our cell count calculations through serial dilutions if necessary as well as to conduct a practice simulation to prepare for the conduction of our very own research experiment through testing the effects that baling wire would have on tetrahymena due to it’s micro plastic composition. The Purpose of these procedures is to help develop our experimental designing and preparation skills.

Procedure:

1. First we must weight a small empty sterile glass jar on a scale and be sure to include all values.
2. We must then take a bundle of baling wire and begin to cut the fibers into very small pieces with a pair of scissors. As you are cutting the baling wire you may want to have the fibers already inside a sterile glass jar.
3. Once you have finished cutting the bailing twine you must weight the glass jar again and record your measurements. To find how much baling twine you have added in grams you must take the weight to the glass jar with fibers in it and subtract the original weight of the empty glass jar. Add or take out baling twine until you have 5 grams of polypropylene in the jar.
4. Once you have 5 grams of polypropylene in jar, you must then take a graduated cylinder and add 50mL of PPT media to it which you will then add to the sterile jar containing your polypropylene fragments and stir.
5. This mixture will then be microwaved for approximately 1-1 1/2 hours at 10 minute intervals of mixing in order to draw out and create a micro plastic solution.
6. Next we will be determining our tetrahymena concentration in a culture by drawing 20μL of the tetrahymena culture and adding it to a Petri dish.
7. Next you will stain the sample by adding 5μL of Iodine to the 20μL drop on the Petri dish and stir with an up down motion by gently taking in and pushing liquid out with the micropipette.
8. A 5μL sample will then then taken from the petri dish and will be added to the center of a concavity slide. You will continue to place 5μL samples one the concavity slide in varying positions until there are 3 total drops of the stained culture on the concavity slide.
9. Observe your findings using the field view objective on the compound microscope and be sure to record all data. Should you need to you may use serial dilutions in order to make cells counting manageable.
10. Now you will draw 20μL of tetrahymena culture and place it on an entirely new concavity slide.
11. Once you add your culture, you will then add 5μL of Indian ink to the the culture and mix with the pipet then add a cover slip.
12. Observe the culture under a microscope and record how many vacuoles you find in 10 cells and record how many vacuoles are present every 10 minutes after until 30 minutes have passed.

Could not reach 30 minutes due to time constraints.

Storage:

Once we finished concluding our experiments we cleaned our workstation and ensured that all materials had been put back in their appropriate location. All microscopes were powered off and stored away. All slides were cleaned and dried. Pipet tips were properly discarded and the micropipettes were also properly stored away.

Future goals:

In conclusion the lab was a successful trial run if you will for our experiments that will be conducted suing very similar methods and materials. In the future I wish that we could possibly take the experiment a step further and conduct an experiment which would determine if micro plastics have a role in the genetic mutation in the DNA of cells.