November 16

Lab 13: Ciliate Isolation and Characterization

Daphne Simo

11/15/18

I. TITLE

Ciliate Isolation and Characterization

II. RATIONALE/PURPOSE

The purpose of this lab was to continue observing soil samples in the non-flooded plates and Falcon tubes in order to find ciliates to classify. Furthermore, the Falcon tubes were analyzed to determine their texture and percent composition of silt, clay, and sand.

III. MATERIALS

Compound Microscope

Concavity slide

Micropipettors

Petri Plate

Soil sample

Dispersing agent

IV. PROCEDURE

Ciliate Observation

  1. Retrieve soil sample from concealed container or Falcon tube from test tube rack
  2. Using a micropipettor, obtain around 10-15μl of the sample and place onto a concavity slide.
  3. View the concavity slide at 4x and 10x magnification to start with a compound microscope.
  4. If ciliate found, obtain images and videos to use for future classification.

Soil Texture Test

  1. Obtain soil sample from storage box.
  2. Remove any debris such as rocks, sticks, plastic, leaves etc. from soil.
  3. Place about 4mL of the debris-free soil into a Falcon tube.
  4. Put about 3mL of water into the tube with soil.
  5. Cover the falcon tube and mix the contents.
  6. Add 1 drop of the dispersing agent. Mix again.
  7. Observe tube after 30 seconds in which the sedimentation of sand particles will have occurred.
  8. Let tube sit overnight or until next lab session.
  9. Use a ruler to measure the percent of sand, silt, and clay. A preferred method is to take a clear picture of the Falcon tube and zooming in on the sedimentation lines.
  10. Find percentage of each type of soil particle.
  11. Record soil type in notebook and on Soil Data Sheet.

V. DATA & OBSERVATIONS

Soil Texture Test

Total of sand, silt, and clay: 2.1 cm

Sand: 1.15 cm (1.15/2.1) = (54.76%)

Silt: .85 cm  (.85/2.1) = (40.48%)

Clay: .1cm (.1/2.1) = (4.76%)

Texture: Sandy loam

Observation of Ciliates

Fortunately, I was able to find ciliates under the compound microscope this week. However, the ciliates were very hyperactive and constantly moved in and out of the different layers of the sample. As a result, I was unable to obtain any good footage of the ciliates.

VII. STORAGE 

The Petri Plates and concavity slides were carefully cleaned off with water, then dried off for future use. The soil samples had their Petri dish lids placed back on and stored in a secure bin. The compound and dissecting microscopes were covered and stored away properly. The Falcon tubes were stored away safely on a tube rack by our lab instructor.

VII. CONCLUSION

I can definitely say that lab this week was better than the previous week. Utilizing the Falcon tube and Bermuda grass as other sources to find ciliates, I was able to finally find some. This was a great step towards progression in hopes to identify a ciliate for the group presentation. However, all of the ciliates I found were either moving too quickly or bobbed in and out of different layers, not allowing me to capture a good image of what I found. Luckily, I have one more opportunity to find ciliates during open lab on Monday. Therefore, I hope to be able to locate more ciliates, and most importantly, capture good footage of them.

VIII. FUTURE STEPS

Moving forward, I strive to become more patient when I am unable to find ciliates. It was very disheartening when I was able to locate a ciliate, but it would move too quickly to be identified, causing me to not get footage. What also deterred me from finding ciliates were the amount of particulates in the samples I viewed. The debris and dirt would move, causing me to lose and ciliate or misidentify one, so I hope to take more clearer and diluted samples.

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November 16

Soil Texture

Isaac Flanagan

Cili Cure

15 November 2018

 

Soil Texture

 

Objective:

The objective of this lab was to be able to find out our soil texture. Another part of this lab was the same as last week’s lab, which was to try to find and isolate a ciliate. After finding and isolating that ciliate, the next step was to stain it, and try to identify it, if you found one.

 

Materials:

  • Falcon tube with separated soil
  • Dissecting Microscope
  • Compound Light Microscope
  • Micro-pipette
  • Soil Sample
  • Ruler
  • Water
  • Cover slide
  • Iodine

 

 

Procedure:

 

Soil Texture:

  • First, get the falcon tube with the separated soil.
  • Take a picture of the tube next to a ruler.
  • Measure clay, silt, and sand in millimeters.

 

Isolation:

  • Get soil sample, and put it under dissecting microscope
  • If you see a ciliate, use micro-pipette to extract it.
  • Put ciliate on cover slide
  • Use compound light microscope to observe it
  • If you can still find it, use iodine to stain the ciliate for better observation

 

Data:

Sand 6mm 35%
Silt 8mm 47%
Clay 3mm 18%

The overall texture came out to be loam.

 

Results:

The results for the soil texture fit the requirements for loam. As for the isolation, I was ultimately unsuccessful. I had a couple of ciliates swimming in my sample, so I extracted them and put them on a slide. After putting them on a slide I used a compound light microscope to observe the ciliate. I found it using the compound light microscope, but when I used the iodine to stain the ciliate I lost it.

 

Conclusion:

Overall the lab was good for teaching us how to calculate the soil texture, which could possibly tell us what kind of soil our ciliates prefer to live in. The isolation part was rather tricky for me, but other than that the lab was pretty fun, as well as helpful.

 

Next Steps:

The next steps are to continue looking for a ciliate, and to be able to identify the ciliate once we find it.

 

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November 16

Lab 13: Soil Ciliate Discovery and Texturing

Objectives:

The objective of this lab was to find a ciliate and capture a picture of it for further identification. We also finished testing our soil classifications by observing the different layers of soil in our falcon tubes.

 

Procedure:

Soil Texture –

  1. Put 4mL of soil from the soil sample into a falcon tube
  2. Add enough water to fill the falcon tube to 10mL
  3. Add 1 drop of dispersing solution to the falcon tube, and shake the tube well to mix the contents
  4. Label your tube with you initials, section number, and the year and place into the test tube rack in the front of the room.
  5. After the contents of the tube have settled over the period of a week, use a ruler to measure the percentage composition of your soil.
  6. Measure the amount of sand, silt, and clay and divide it by the overall measurement of soil.
  7. Record results in your lab notebook.

Ciliate Discovery –

  1. Use a clean slide to scrape your soil onto one side of your petri dish
  2. Let the water drain from the soil and collect on the opposite side of your dish
  3. Use a pipette to place many drops onto a slide for observation.
  4. Observe each drop and search for any organisms
  5. Take pictures and describe any organisms found

 

*group picture of single ciliate*

 

Data:

Soil Texture –

Total Amount of Soil: 1.6 cm

0.7cm – sand     0.7/1.6 = 43.75%

0.8cm – silt        0.8/1.6 = 50%

0.1cm – clay       0.1/1.6 = 6.25%

Soil Composition Name: Silt Loam

Ciliate Discovery –

Only one ciliate was discovered that was able to be properly photographed, and it is pictured above. Another organism that was found were Nematodes, which were moving around erratically like in past observations.

Storage:

All lab equipment was stored properly, and microscopes were covered and placed in the middle of the tables for future use. Our samples were returned for future use and observation, and out tables were cleaned.

Conclusion:

We have yet to identify our ciliate, but we do have a picture that will help to compare it to other known species of ciliate to further identify it. Nematodes continue to be found all throughout the soil samples and could be the reason ciliates seem to be limited.

Future Goals:

In the future, I would like to repeat the experiment with a better sample of soil to hopefully capture and identify more ciliates. I would also like to get a higher quality picture of my ciliate along with a video showing its movement pattern.

 

 

 

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