Aynsley Gaspard

9-27-18

Biology Lab                                                   Ciliate Challenge 6

Purpose:

The purpose of this lab was to enhance our already acquired laboratory skills, like pipetting, serial dilution and cell counting. We also were introduced  to micro plastics. We were to learn about how to make “twine juice” using hay-bailing twine. We will let them sifter a week, then test their effects in soil ciliates.

Procedure:

Polypropylene Challenge

1. Shred the polypropylene (black) twine into small pieces.
2. Measure 0.5 g of twine into sterile glass.
3. Add 50 ml of sterile PPT media.
4. Boil and stir for entire lab.
5. Filter 5 ul filter paper in sterile into 50 ul tube.
6. Autoclave “twine juice” and store for a week.

Serial Dilutions

1. Add 20 ul of cells to 5ul of iodine to Petri Dish
2. Mix up and down by pipetting.
3. Add 3 5 ul drops to a slide and count stained cells; record average
4. To dilute 1:10 on petri dish , add 5 ul of first drop to 45 ul of PPT media
5. Record your counts and report average in cells/ml

Directional Movement Assay

1. Add 5 ul of drop of culture to a clean flat microscope side.
2. Place slide one dissecting scope.
3. Focus slide on black plate using reflecting light.
4. Choose a cell to follow for 10 seconds.
5. Keep track of how many times the cell changes direction more than 17 percent during this time period.
6. Record number of direction changes.
7. Repeat for at least 10 cells.
8. Record the numbers and calculate the average number of direction changes per 10 seconds.
9. Calculate Standard Deviation

Cells                                Number of Directional

Changes/10 sec.

Average Standard Deviation: 1.032 number of directional changes/ 10 sec.

Conclusion:

We learned how to challenge our knowledge of ciliates and furthered our knowledge a little but moron micro plastics. We are now able to begin testing the effects of micro plastics on tetrahymena. This will contribute to our experiment our group is creating.