Objective: Today we learned how to find the T test, F test, histogram, and descriptive analysis of data using Excel. I have attached the data for the 1 microliter group. Based on the P-values calculated, I reject the null hypothesis.
T-test of control and treatment, P-test for control and treatment
Descriptive statistics for control and treatment
F-Test and P-value between treatment and control
Histogram of control
The purpose of this lab was to test the experiment we created. The purpose of the 31 lab, was to test the effects (if any) that insulin has on tetrahymena. First we practiced more serial dilutions and practiced with calculating the number of cells in a solution. Next, we determined that the dilution my group (A) would be using, is .1 ml concentration of stock. 999 microliters of PPT (protein base) and tetrahymena were added to 6 well plates. 1 microliter of water was added to 3 control groups. 1 microliter of insulin was added to 3 separate well plates. 10 microliters of each well plate were placed on a concave slide, and we used the compound microscope to count the number of tetrahymena. See picture for the result of tetrahymena counted. On day 2, I added 20 microliters of iodine to each well. I mixed the wells and added 3 separate drops of 10 microliters to a concave slide. I counted each drop and recorded the number (for more info, see picture below) of tetrahymena. Next I found the average of the 3 drops and recorded that number in the excel spread sheet. The general conclusion I would draw from this lab, is that insulin increases the population of tetrahymena by a significant amount. I was not expecting there to be so many tetrahymena, so I was surprised that the number increased so rapidly. Some errors I encountered during this experiment was, I initially added three drops of 10 microliters to one portion of the concave slide, causing my numbers to be really high. Also, it is a possibility that I miscounted the number of tetrahymena, which affects the averages. My cell well plate is stored in the bottom drawer with my name and section 31 labelled on it.
The purpose of the Ciliate Count Challenge lab, was to practice counting tetrahymena in a sample and to practice diluting solutions. The first step was to take a 10 microliter sample of tetrahymena, put it on a concave slide, and count the number of tetrahymena observed under the compound microscope. The number of tetrahymena was too many to count, but I guessed that the number was around 200. Next I took 90 microliters of PPT (protein culture) and mixed that with 10 microliters of tetrahymena. I added 10 microliters of this solution to a (new) concave slide and counted the number of tetrahymena using the compound microscope. At first I accidentally counted only one section of the concave slide, and counted 9 tetrahymena. I recounted the number three more times and got 21, 24, and 24. Next I found the concentration of the tetrahymena (see calculations in picture). To find the salt concentration, I added 10 microliters of NaCl to 90 microliters of water (and mixed them together). I then added 10 microliters of this solution to 10 microliters of tetrahymena, and all of the tetrahymena died. I then added 5 microliters to 90 microliters of water (and mixed them together) and added 5 microliters to the tetrahymena (10 microliters). Some of the tetrahymena died, but not all. Next I added 20 microliters of NaCl to 80 microliters of water. When I added 10 microliters of this solution to the tetrahymena, there was no visible change in the tetrahymena. Next my group calculated the concentration (see picture). In total, there were 24,000 cells/ml and the NaCl concentration was 125mM. We will be using the averages from the cell counts and compare them to rest of the class. The average cell counts will be applied to our group experiment next week. The next step, is to perform our experiment. We will be studying the effect insulin has on tetrahymena. We did not store our samples for this experiment.
In today’s Serial Solution lab we learned about micropipetting and how to properly use or handle micropipettors. The purpose of this lab was to practice using pipettes and making dilute structures. The purpose of this lab was to teach us how to calculate the concentration of a solution mathematically, for both diluted and undiluted solutions. We used media and tetrahymena culture to make a 1:10 dilution in tubes. First we took 90 microliters of PPT-which is a protein media for tetrahymena- and placed that into 4 tubes. Next we added 20 microliters using the micropipettors to each of the 4 tubes. After that, we spun the tubes to make sure the solution was properly mixed together. Then I took 10 microliters from the first solution using a micropipettor and placed it into the 2nd solution. Next I took 10 microliters from the 2nd solution and placed it into the 3rd solution. After that, I took 10 microliters from the 3rd solution and placed it into the 4th and final solution. I proceeded to place 5 microliters from each tube onto a concave slide. Next I tried to count each tetrahymena in each sample using a compound microscope under 40X but that did not work out. Instead, I took 5 microliters of undiluted tetrahymena and counted the number. Unfortunately, there were too many to count so I had to estimate. I estimated that there were about 80 tetrahymena. From there, I proceeded to calculate the average cells for each dilution (see chart in picture). the overall interpretation is that the number of tetrahymena decreased as dilution increased. I made several errors throughout this experiment that could have affected my results. It has just occurred to me that I did not mix the sample tubes after I diluted them with an additional 10 microliters of a solution. Also I placed all of the samples on one slide (due to time restraints), which made it extremely difficult to see the tetrahymena because I had to keep readjusting the stage. Next time I will only put one sample at a time on the slides. (we did not store our samples)
The purpose of this lab was to practice and learn how to read primary articles. Learning what to look for in primary articles is important because they can aid in personal research and studies. Primary articles give a detailed description of an experiment and they are often written by the scientists who created the experiment. The Primary article I read was Biodiversity Patterns Of Soil Ciliates Along Salinity Gradients. The central question investigated was: what are the affects of salinity on soil ciliates? This experiment revealed that soil salinity and water are the main factors that regulate the distribution of ciliates. Six different coastlines were chosen with different salinities for this experiment. The types and population of ciliates were measured at each location. Seven ciliates were found in total. Sequences of genetic material were also taken from each type of ciliate. The result was, that there is a higher diversity of ciliates in non saline soil. 8.9 psu (as indicated by table 2) had the highest amount of diversity and population of ciliates. The locations with the highest amounts of salinity had the lowest amount of ciliates, and the lowest diversity. This was deemed to be a surprising result because ciliates do prey on bacteria, and bacteria live in environments with a high level of salinity. It was predicted the ciliates would thrive in the highly salinity envirnoments because they would have abundant prey, but the opposite occurred. Despite the fact that the high salinity areas also had a high water content, the ciliates still did not survive in that environment. This therefore proves that salinity is a major environmental determinant, not water. The main question I had throughout this article was: why was it necessary to study the rRNA of the ciliates? Tables 2 noted that rRNA was sequenced, but I do not understand the reason why that was done. Over all I do agree with the conclusion that salinity plays a major role in soil health and the diversity of ciliates (and protists in general). It is important to keep a stable level of salinity in order to preserve the microorganisms living there. Since I have discovered that salt plays a major role in the biodiversity of ciliates, I would like to now investigate the different affects that sugars (such as cellulose and starch) have on soil ciliates. Here is a link to the article: http://s-cdn.com/S0932473915001066/1-s2.0-S0932473915001066-main.pdf?_tid=664d44a8-995a-11e7-af41-00000aacb35f&acdnat=1505400100_65ec50ec6d392bccfc128d19866d526eac.el
The purpose of this lab was to practice or learn how to use a compound microscope. Last week we used the dissecting microscope to view the ciliates, and this week (9/07/17) we used the compound microscope to view the ciliates. We used the compound microscope this week in order to view the ciliates on a more magnified level and to get a better, or more detailed picture of what the ciliates look like. In this activity, each person was assigned two cultures. I was in charge of documenting cultures A and B. We had to transfer a small drop of culture using the pipettes onto a concave slide. Then we had to observe the specimens under the 4x and 10x magnifications. I had a difficult time observing both samples under the 10x magnification. I was able to get a really detailed view of my sample B ciliate. This ciliate is fairly long and I could see the organelles inside of the ciliate. I think this would represent a blepharisma or stentor ciliate. I think it could be the stentor ciliate because, under the 10x magnification, I was able to see what looked like a hole or opening similar to the one stentor ciliates have. There are some elements of the shape of ciliate B however, that leads me to believe it could be a blepharisma. Ciliate A looks like a paramecium. It had the visible cilia, it was really tiny, and it moved quickly. This experiment was important because it allowed us to practice using the compound microscope properly and we were able to practice identifying more ciliates. My sample of ciliates were bleached so they are currently not alive anymore (location is no longer in the drawer).
The purpose of this lab was to practice identifying different types of ciliates that can be found. The goal is for us to learn how to recognize different types of ciliates, so we can properly identify them in our soil samples later on. We took 6 unknown samples and placed them into a 24 cell well plate. Next we placed each sample under the microscope and viewed what each one looked like. Next we had to sketch a picture of what we saw and document some key characteristics, such as size, shape, movement, and location. The 1st sample had ciliates that looked like tadpoles. They moved quickly and were located in the middle of the media. The 2nd sample had ciliates that were long and slender. These ciliates were primarily clustered at the top of the medium. The 3rd ciliate looked like a tiny sausage. It was wide and long, and it rotated while it moved in different directions. The 4th sample was shaped like a blob. It moved very quickly. I was able to get a very detailed view of what the inside of this ciliate looked like (I was able to see the 2 nuclei). The 5th ciliate was an oval shape. I was able to get a great view of this ciliate as well. I saw the cilia moving and propelling the ciliate around. The 6th ciliate looked like a worm. I had a difficult time getting a good picture of this ciliate but I noticed that it was very slender and moved slowly. I have attached a picture of the sketches I drew, and I stored my samples in the top drawer near my seat. It is labelled LNM31Fall2017.
A: Stenor, B: Blepharisma, C: Paramecium, D: Euplotes, E: Paramecium, F: Spirostomum
I collected a soil sample for this lab to investigate the different organisms that are in soil. The end goal, is for us to study the types of ciliates (or maybe discover new ciliates) in soil. I collected my soil near the Marrs Mclean Science Building. I choose soil in a shady spot a couple feet from a large tree. The soil did have a lot of acorns in them so I had to pick those out of my soil sample. The exact location noted on my GPS is Bear park, 1200 S 4th Street. The time I took this soil sample was at 11:41 Am, the temperature was 91 degrees, and the humidity was 48%. The last rainfall at the time was 8/17/2017. In class we put a portion of our soil sample in a petri dish and found the mass of our samples. My soil sample weighed 39.0g (including the weight of the petri dish). We have yet to discover what types of ciliates or microorganisms are in our soils. My soil sample is labelled with my initials (LNM), section number (31), and fall 2017. It is located in the top drawer near my seat.
