Objective of this lab:

There were multiple objectives for this lab. The first 2 objectives were to be able to understand how a compound microscope works and to understand where ciliates are on the tree of life. After this lab we should also be able to make a wet prep and focus using 4x, 10x and 40x and be able to use methyl cellulose and stains to better observe the specimens.

Purpose of this lab:

The purpose of this lab was to correctly utilize a compound microscope so that we are able to see the ciliates in more detail. In addition we will be able to use our Field of View to estimate the size of our specimens.

Procedure:

1. Uncover compound microscope and using a plate with Lycopodium Strobilus, go over how to focus the microscope correctly. Doing this we make sure to focus using the coarse adjustment only on 4x.
2. Next we use the millimeter side of a ruler to measure our field of view and we use the equation FOV (low) x Mag (low) = FOV (high) x Mag (high). We are also sure to convert to micrometers.
3. Using a dissecting scope we choose one of six unknown specimens and place a drop onto a concave plate  so the ciliates are able to move around. This is the wet mount.
4. Place wet mount under compound microscope and focus it. Record data. Recording should include measurement of specimen, color, movement and pictures.
5. Do the same for 10x and 40x.
6. Remove wet mount and set aside concave plate.
7. Using a flat plate take a drop of same specimen and add a drop of methyl celluose. Record data at 4x, 10x and 40x again.
8. Using a flat plate take 2 drops of specimen. Place one drop on each side of the plate. On one drop place methyl green pronin Y. On the other drop place Lugol’s Iodine. Record date at 4x, 10x and 40x from both drops.
9. Clean off plates with bleach in sink and leave them to dry.
10. Store compound scope properly.

Observations:

Observing my specimen on the wet mount at 4x magnification, I found the specimens were small and had a reddish tint to them. With my field of view at a diameter of 4mm (diameter in micrometers was 4000) I estimated around 20 ciliates could fit across. There was a 200 measurement. At 10x the diameter for my field of view was 1.6 mm (1600 micrometers). The measurement was 80. Still using the wet mount my specimen at 10x still had a reddish tint. Through 10x I was able to see the patterns of the ciliates more clear. Some ciliates seemed to be moving and other didn’t. If they were moving they were very slow. At 40x my field of view was .4mm (400 micrometers). The measurement was 20. This was the last magnification using the wet mount. In 4ox only one ciliate was seen and it was moving so slow it looked as if it was barely moving.                                    Next I used a flat plate and added a drop of methyl cellulose. Methyl cellulose is supposed to slow down the ciliates. My first drops of specimen and methyl cellulose did not pick up anything. I re-tried and could only find one organism. The ciliate was not moving at all and the measurement was 200. At 10x and 40x I could not pick up on anything other than the one ciliate. It was still not moving. Next using a flate plate I put one drop of specimen and one drop of Methyl green Pronin Y. This is supposed to stain the ciliates. With this you have to observe quickly because the cells will begin to die off. At 4x for this one I could identify one ciliate. It was not moving at all. At 10x I could see the same ciliate in more detail. At this point it appeared a tinted color because of the Methyl Pronin Y. At 40x the ciliate was much more clear and more details became available. You could somewhat see the texture of the outer part of the ciliate as well as parts of the inside. Some organelles also became into view although it was not completely clear. Next I used Lugol’s Iodine. This is supposed to shrink the cell size. At 4x I identified one ciliate and the Iodine caused it to have a orange-yellow tint. The ciliate was around 2 mm long (2000 micrometers). The ciliate was completely still. At 10x the ciliate was still completely still but the image was more clear. The color of the dye tinted the color of the ciliate making it less opaque. At 40x I could not locate the ciliate and the glass was tinted yellow.

Wet Mount

Methyl Green Pronin Y

 Methyl Cellulose

Storage: Glass Plates are washed with bleach and left out to dry. The compound microscope is unplugged and the cover is put back on it.

Future goals: In the future labs being able to focus the microscope faster will help greatly. Something else that will be helpful is being able to identify some ciliates using the compound microscope.

Objective of this lab:

The objective of this lab is to ensure that we are aware of how to use a dissecting microscope. By knowing this we can apply it to being able to clearly see ciliates through the scope.

Purpose of this lab:

The purpose of this lab is to view different ciliates through the dissecting scope and be able to identify what type they may be based on characteristics, shape, size movement and multiple other factors. In doing this we will gain knowledge on how to properly work the microscope and on the process of identifying ciliates.

Procedure:

1. Uncover Dissecting Scopes and go over parts of microscope.
2. Clean 24 well plate; Do this using a plastic pipette (each unknown goes in a separate plate no more than 1/2 full).
3. There are 6 unknowns per groups of 3.
4. Review 6 different unknowns and describe their shape, size, movement, location in media, characteristics and draw a sketch of the ciliate.
5. Be sure to clean up your station. This includes cleaning the plates with bleach, cleaning the microscope and wiping the table.

Observations:

The six different unknowns all had unique characteristics that set them apart. This was helpful in identifying them. At first it was difficult learning how to focus the microscope in a way where you could clearly see the unknown. This was partly because the different unknowns floated at different levels in the liquid making it a constant point to re focus the microscope. Unknown 1 was oval like and relatively small. It moved somewhat quicker than the others and was located all over the media. It was clear with black points on the end and had a tadpole like shape. I identified it as possibly a paramecium. For this one the focus was at 2.6 Unknown 2 was circle like and small. It moved slowly in circles while also spinning. There was no specific location in the media, it was spread out. It was clear with a string like body and was also flat like. I identified it as possibly Aspidisca. For this one the focus was at 3.4. Unknown 3 was long and oval like and somewhat large compared to the others. It moved slowly and was spread out all over the media. It was long, dark colored and not as transparent as the others. I identified it as possibly Frontonia. For this one the focus was at 4. Unknown 4 was also oval like and long. It was small, moved in slow circles and was located all over the media. This unknown had a red tint to it. Identified it as possibly Spirostomum. For this unknown the focus was at 4. Unknown 5 was round and mostly in the center of the media. It was extremely small, black and moved slowly in circles. I identified this one as possibly being Euplotes. For this unknown the focus was at 3.5. The last unknown (6) was oval shaped and not flat. It was not small or large and did not move fast. It was free swimming and its presence was all over the media and it had a bluish tint. I identified this one as possibly being  Stentor.

Storage:

We dispose of the ciliates in the sink and clean the small plates with bleach. After this we left the plates on the left of the sink, right side down , to dry.

Future Goals:

In the future I want to be able to focus the microscope quicker. This will give me more time to be able to observe the unknown ciliates.