Lab 12: Ciliate Classification 11/9/17
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-
I saw a couple nematodes within my original soil sample and there was some rapid movement from very small organisms. I think that these very small fast organisms swimming in the water were ciliates. There were about 3-4 ciliates present but they were so small it was difficult to tell/ It was hard to focus on them under the compound microscope because they were transparent and round and could easily have been mistaken as tiny particles of dirt. As I viewed the sample using a higher power under the microscope, it became more clear that they were ciliates and they were so transparent that they were difficult to differentiate from the water they had been swimming in.
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:
In the future I hope to be able to actually identify the ciliates that I discovered in my soil sample. I would also like to be able to classify them as well as know what type of ciliate group they belong to and where they would be located within the ciliophora subsection of the phylogenetic tree. I hope that I will be able to conduct more research regarding ciliates in the future and eventually understand how they contribute to our ecosystem.
