Objective:

Today our goal is to finally conduct our experiment by testing whether of not the concentration of insulin that tetrahymena are exposed to affects their growth or death rates. We will perform serial dilutions with a control group of water and a treatment group of insulin to conduct the experiment. Each group will be assigned a dilution factor in which each group member will test three times to maximize our trials per dilution factor in the class as a whole.

Purpose:

The purpose of conducting this experiment is to finally test how insulin affects tetrahymena, specifically how it affects their growth and/or death rates. The purpose of this experiment as a whole is to continue practicing serial dilutions so that in the future we will be able to conduct our own research.

Procedure:

1. Obtain a sterile 12 well vwr tissue culture plate, an insulin stock solution, sterile water, a tube of standard tetrahymena media, various micropipettes and their corresponding tips, as well as concavity slides.

2. Assign three wells to be the control group and three wells to be the treatment group. There are three wells of each to test the designated solution factor three times for both water and insulin.

3. Use a P1000 micropipette to take out 990 μl of the tetrahymena media solution and add it to all six wells that will be used for this experiment.

4. Use a P20 or a P10 micropipette to take out 10 μl of sterile water and add it to each well of the control group (using a total of 30 μl of water).

5. Use a P20 or a P10 micropipette to take out 10 μl of the insulin stock solution and add it to each well of the treatment group (using a total of 30 μl of the insulin stock solution).

6. To ensure the samples are mixed thoroughly, it is possible that they may need to be stirred using a micropipette tip.

7. Using 990 μl the tetrahymena media with either 10 μl of water of 10 μl of insulin creates a 1:10 dilution factor.

8. View a 10 μl drop from each well and count how many ciliates are visible and record your observations.

9. After 24 hours (open lab) view another 10 μl drop from each well sample and count how many ciliates are visible and record your observations.

10. Since you used a 10 μl sample, you must the number of dilates counted by ten, then you would multiply the amount of ciliates by 1000 to account for the 1:10 dilution factor, and this will tell you how many ciliates were present in each well sample (approximately).

11. Compare data from the initial number of ciliates counted for each sample to the number of ciliates counted after 24 hours.

Data and Observations:

After comparing the number of ciliates counted between the two samples, I know that the insulin has had to have made the tetrahymena able to produce faster as they absorb more nutrients. I think that the 1:10 dilution factor was a good amount because it did not seem like too much because they did not die within 24 hours. They thrived more in the insulin than in the control group containing water.

Current Storage Situation:

The well plates were labeled and place in the bottom middle drawer on the front side of the desk, facing the projector, containing member of groups C and D. Mine is personally in the far bottom left corner within the drawer nearest to walls of the drawer. It is labeled KSA31F17.

Future Goal:

After the experiment is finished I hope to be able to use my data and apply statistical analytic programs used in Excel in order calculate things such as standard deviation, mean, and t-Tests as well as F-Tests. I hope to gain further knowledge of how to successfully use this software so that I can apply it to any researchI conduct in the future.