9/27/18

Objectives/Goals: This particular lab had many objectives; creating the microplastic solution, practicing a cell assay, and refining our cell-counting techniques. The overall goal was to become proficient at efficiently getting our cell counts and becoming familiar with performing different assays on the cells.

Materials: Petri dish lid, micropipettors, Tetrahymena stock solution, flat slide, dissecting microscope, compound microscope, iodine, ruler, baling twine, scissors, scale.

Procedure:

PP Microplastic Production-

1. Cut the polypropylene (baling twine) into small pieces with scissors
2. Measure 0.5 g of the PP in a sterile glass jar
3. Boil (not done in class)
4. Filter with 5 μl paper (not done in class)
5. Autoclave and store (not done in class)

Ciliate Count Challenge-

1. Add 20 μl of TH to 5 μl of Iodine on a petri lid
2. Mix
3. Add 3 separate drops (5 μl) to a slide without coverslip
4.  Count cells using 4x or 10x objective
5. Record all 3 trials

Simple Swim Speed Assay-

1. Place 20 μl drop of culture on flat slide
2. Set slide on top of ruler between 2 mm lines (1 mm gap)
3. Pick a cell to watch
4. Time how quickly the cell can swim 1 mm
5. Record for 10 different cells
6. Calculate average and standard deviation

Data:

Observations: The cells in the 3 trials were simple to count. I easily observed the swimming speeds of the tetrahymena with the video I took (I used Dr. Adair’s phone lens apparatus).

Storage: The slides were rinsed with the cleaning solution and water and were placed on paper towels to dry. The microscopes had their cables wrapped and were covered. The used micropipettor tips were discarded, and the micropipettors were hung on their racks. The stock solution, Tetrahymena media, and well plates were left on the tables for the instructors to handle.

Conclusion/Goals: This lab went efficiently, and with no mishaps. I was able to perform all 3 trials for the cell count and all 10 trials for the swimming speed. The video I took of the tetrahymena swimming was very helpful because I was able to slow the video down and note the exact times. Overall, I am pleased with how this lab went. I am able to tell that with each lab, I am becoming more skilled at the lab techniques. My next goal is to become more precise when counting the tetrahymena.