Objective:

The goal of today’s lab was to see if we were able to view any ciliates from the new culture of ciliates. The goal of today was to mainly observe whether or not we had successfully isolated ciliates from our non-flooded plates and if they had effectively been cultured. If we were not able to see any ciliates we had to go back to our non-flooded plates and repeat the process stated in lab 12.

Purpose:

The purpose of lab was to be able to view ciliates that we had isolated from our soil and possibly identify/ classify them. This would be helpful practice for identifying ciliates or even trying to identify some specific features of the ones that were able to be observed under the microscope.

Procedure:

1.) Retrieve the well plate with cultured ciliates.

2.) View the whole well under a dissecting microscope to see if any ciliates are present within the well.

3.) If ciliates were present within the well, pipet ten microliters onto a concavity slide and view under a compound microscope.

4.) If there were no ciliates present within the well, retrieve the non-flooded plate and try to isolate and extract a ciliate from that sample.

5.) Once a ciliate is able to be seen under a microscope, try to classify and and identify the ciliates present.

6.) Try to capture an image of any of the ciliates that can be seen under the micro scope.
Note: Some ciliates may be too small to identify using the compound microscope.

7.) These images should be used during your group presentation of your findings during this experiment.

Data/Observations:

I observed a few ciliates that were too small to identify and could best be seen using 100x on the compound microscope. The ciliates I had seen were more oval shaped with very distinct organelles within due to its transparent body.

Current Storage:

My non-flooded plate and 24 well- plate are labeled with my soil identifier, KSA31F17, and are located in the left back desk in you are facing the projector in the second drawer on the front side of drawers closest to the next lab table. The falcon tube that was used to calculate the soil texture in located back in the teal- green tube stand with the other students tubes on the back lab counter closest to Dr. Adair desk top computer.

Future Goal:

I hope to be able to collect more soil samples in the future and be able to identify more ciliates or at least classify the group that the ciliate belongs to. I hope to use my knowledge I gained from this lab for any future research career that I choose to pursue.

Objective:

The objective of this lab is to apply core concepts of evolutions, classifying ciliates using phylogenetic trees as well as determine the soil texture of our samples. The main goal of this specific lab is to eventually improve these phylogenetic trees and our understanding of the diversity of life within the ecosystem.

Purpose:

The purpose of lab today is to observe and understand how ciliate populations evolve and how the change in genetic composition effects the population over time. We hope to eventually see how populations evolve when individuals with different genotypes survive and reproduce at different rates.

Procedure:

1.) Retrieve your falcon tube from the teal-green rack and use a ruler to measure the amounts of sand, silt, and clay within the tube because everything has settled by now. Measure the total amount of sand, silt, and clay as well as take individual measurements of each ()this will help to calculate soil texture).

2.) Calculate soil texture by dividing the individual measurements of sand, silt, and clay by the total measurement of all three of them and then multiply that number by 100 to find the actual percentage.

3.) Retrieve the non- flooded plates created from the previous lab and observe them under a dissecting microscope. Use the microscope to see if there are ciliates resent within the water of the soil sample. Try your hardest to find anything that may resemble a ciliate and if you are unsure ask Hope, Felicia or Dr. Adair to check.

4.) Once you have a found a ciliate or multiple ciliates, using a p10 to micropipette ten micro-liters of the soil sample where you viewed the ciliate on to a concavity slide. It may take multiple tries to successfully capture the ciliate viewed on the microscope into the ten micro-liter extraction.

5.) On the concavity slide with the ten micro-liter drop containing the ciliate from your sample, use a compound microscope to ensure that you in fact have a ciliate within that sample and that it is visible under this microscope.

6.) Once you know that you for sure have atleast one ciliate within your ten micro-liter sample on the concavity slide, obtain a twenty-four well plate.

7.) In one well of the well- plate, pipet five hundred micro-liters of the protozoa pellet solution and then add as much of the ten micro-liters from your concavity slide as you can into the same well on the well- plate. This will help to culture any ciliates present in your sample.

Data/ Observations:

Soil Texture-

Objective:

The objective of today’s lab was to measure soil characteristics as well as ciliate diversity and abundance. Our goal is to understand where and how ciliates get their nutrients and to possibly identify what environments make it possible for them to thrive.

Purpose:

The purpose of lab is to be able to calculate soil water content, create non-flooded plates and determine the pH of the soil. After learning how ciliates function and knowing what environments have the most diverse groups of ciliates we will be able to possibly understand their true functions and why they are a vital part of the ecosystem.

Procedure:

1.) Retrieve your original soil sample that was collected during the first week of class from the drawer the that it is located in (if you do not remember where you put it, check the first entries of your lab journal and should be noted somewhere).

2.) Weigh your sample on a balance and record the data. Look back to the first lab and find the original weight of your sample and calculate the percent of water. You can do this by subtracting the dry soil from the original weight of the wet soil and dividing that number by the wet soil. Then record what you got in your lab journal and add it to the class data sheet.

3.) Create a non- flooded plate and saturated the soil with sterile water. There should be just enough water that when you slant the plate a small pool collects to one side rather than there being so much water the entire plate is just a pool of water.

4.) Observe the non- flooded plate using the dissecting microscope and look to the edge of the water that has pooled together for ciliates. It is common to not see any ciliates but make sure to record all initial observations made in your lab journal and remember that excystment takes place within 24-48 hours and more ciliates maybe be seen after that time period.

5.) Obtain a falcon tube and fill the first 3mL with the soil from your sample and then fill with water to the 8mL marker. Label this falcon tube with your initial. course number, and semester year (KSA31F17). After you have a filled falcon tube, shake it and the let it sit so that the sand, silt, and clay can settle and the soil texture can be calculated.

6.) After the soil from the falcon tube has settled for a little bit, pipet 1mL into a micro centrifuge tube and put it in the centrifuge for one minute so that are the sold particles within the water create a pellet at the bottom of the tube.

7.) Pipet out only the liquid carefully so none of the pellet from the bottom is obtained. Pipet the liquid into a small glass bottle.

8.) Obtain pH paper and place a small amount of it and place it in the small glass bottle with the water from your sample. Shake the glass bottle until the pH changes color. Review the pH color wheel to determine the pH of your sample and record it in your lab journal.

9.) Return back to your falcon tube that has been sitting and add one drop of the soil texture into the tube and shake once more. Place the tube in the green rack on the counter and let it sit until next lab, this will enable you to actually calculate the soil texture during the next lab.

Data/Observations:

Mass of Empty Petri Dish- 5.7g
Mass of Petri Dish with Wet Soil- 35.5g
Mass of Wet Soil- 29.8g
Mass of Petri Dish and Dry Soil- 28.0g
Mass of Dry Soil- 1.8g
Percent of Water Content- 6.04%
pH- 6.5

My soil was very dry and needed a lot more water to even create the non- flooded plate compared to of individuals. The soil itself was very mulch like and had a lot of clumps, twigs, and even some small branches. The texture was very thick and it was a very dark color that resembled more of a black than a brown.

Current Storage:

My falcon tube is currently stored on the back lab counter within a teal- green stand with the other students falcon tubes and is label with my soil identifier, KSA31F17. The non- flooded plate is located in the same drawer it had been in which is located on the back left table on the second drawer on the front side facing the projector.

Future Goal:

After completing this experiment, our eventual goal will be to identify and extract ciliates from our soil sample and culture them. Following all of these procedures will also help us for any future research especially if that research is related to ciliates in any way. Using all the equipment and properly measuring things will help us create a foundation for any later laboratory research we may participate in.

Objective:

Today, the goal of lab was to present our final figures and receive advice about how to make the best figure to put in our lab reports. After the presentations the majority of class was spent on finalizing our rough drafts. The main goal of class today was to be able to ask questions and find out how to structure these lab reports.

Purpose:

The purpose of today was to prepare us for our final presentation at the end of the course as well as for submitting our final revision of our lab reports. Today gave much insight into the future and how we as a students will have to perform when presenting scientific information whether that may be within a report or an oral presentation.

Procedure:

1. Previously have finished a draft of the lab report with most to all sections completed.
2. Present the final draft of the group figure as well as record information from other groups which may be helpful to add it to the “Discussion” section of the report.
3. Work with your group to make any revisions to the final figure after the presentation.
4. Ask questions about Lab Report to ensure it is structured in the best way possible.
5. Finalize rough draft and submit it by the end of the class period.
6. Continue to work this draft before meeting with Felecia one on one.

Data and Observations:

Observing all of the other groups presentations helped my group in realizing what we ourselves needed to fix based on Dr. Adair’s comments. Our presentation was not perfect and we needed to correct moles per milliliter to micro moles per milliliter. Many students were struggling in similar ways as I am so listening in to the advice they were being given was very helpful to me when revising my own draft. There is no real quantitative data for this lab but everyone was working very hard to try and understand what they needed to do in order to finalize their drafts as well.

Future Goal:

After completing this rough draft, my goal is to continue to revise it and make it more presentable when I meet one on one with Felecia. I also hope to be able to write better more official lab reports in the future. Learning how to write these reports now will better help as a further my education at Baylor as well as any future research I participate in.

Objective:

The goal of lab today was to learn how to correctly create a figure using excel. Student needed practice using the Excel graphs and how to manipulate them in a way that they portray all the information that is necessary in order for individuals to interpret the information presented. The main objective of today was to learn how to use Excel better and more efficiently when interpreting data.

Purpose:

The purpose of today’s lab was to ensure that we are prepared to write our research papers with figures that have the correct information and are formatted in the best way. This lab is very important, not only for this experiment and course but for any future participation in research. Knowing how to use Excel will help to interpret data to create even more data that could lead to a researcher knowing whether or not they are able to reject or fail to reject their null hypothesis.

Procedure:

1.) Open the Excel spreadsheet containing all of the class data.
2.) Record all of the data pertaining to the students who were testing the same dilution factor as your group.
3.) Open a new blank excel sheet and label two adjacent columns “treatment” and “control”. Then record all of the data from each trial into their corresponding columns on this new spreadsheet.
4.) Once all of the data is recorded, take the average of each one.
*The average is necessary in order to create a wholistic view of the data collected.
5.) Within Excel, click “Insert” located on the top and in the drop down menu, click “recommended charts” which will give a variety of graphs that interpret in different ways. Choose the bar graph because it is the best way to interpret this data.
6.) After selecting the bar graph, move the cursor over to the chart and click it. Then in the top right corner select “add chart element” and scroll down and select to add error bars.
*The information that is needed for this step was calculated in the F-test as standard area for two variables.
7.) Within the “error bars” tab, select “more options” then insert the standard error manually based on the descriptive statistics calculated from the previous lab. This process will place error bars on the two variables shown on the bar graph.
8.) In order to change the color and style of the bar graph, double click on the bar graph and you can select “add chart element again” and from their you can add both vertical and horizontal axis titles.
9.) Label the vertical axis “cell count of tetrahymena (cells/mL)” and then label the horizontal axis “treatments” (label the individual bars “control or water” and “insulin” according the corresponding data presented).
10.) In the top left corner next to the “add chart element”, there is an option to select “quick layout”, if you select this option, then click the seventh layout format from the top and will automatically provide space for a title, and a space to label both axis.
11.) To complete the figure, save the chart as an image (jpeg) and place it in a word document to add a figure caption, then save the whole document and your figure is complete.

Data/Observations:

Current Storage:

During this lab and a few previous labs, we have been working in the computer lab and there are currently no trials or experiments being conducted at this time which would mean “current storage” would not be applicable while as we are writing drafts for our research papers.

Future Goal:

This lab will definitely help us for conducting research in the future because learning how to properly document our findings and results will ensure that others can read our reports and be able to recreate this experiment for future questions that arise regarding ciliates intake of insulin and how it affects them. Learning how to use excel will definitely ensure that we are future researchers will be proficient in not only conducting research but documenting our findings properly.

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.