BIOL 1115 Human Biology Lab 13– Immunology

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13

INTRODUCTION 

Go to http://www.hhmi.org/biointeractive/immunology-virtual-lab

Start the Virtual Lab and maximize the screen if you wish. Answer the following questions

DIAGNOSIS 

1. Where are antibodies found?

2. How can they be used in the laboratory?

3. What does ELISA stand for?

4. What are ELISA assays used for in labs?

5. What are the three important limitations of an ELISA? Explain each.

BACKGROUND 

1. What test can be used to determine whether a patient has an infectious or autoimmune disease?

2. What does a positive result indicate?

3. The watery fluid of the blood is called ____________________.

4. What is allowed to react with the target antigen?

5. Detection is possible when _________________________________________________.

6. Once isolated, the secondary antibody can be ____________________________________

7. What is the signaling system?

8. What happens when the appropriate chemical (substrate) is added?

9. How is the test quantified?

10. What does the amount of color reflect?

LAB NOTEBOOK 

Proceed through the entire lab simulation protocol. Be sure to read the captions below the pictures (left side) and the information in the lab notebook (right side). Be sure to “start over” to begin the lab. You CANNOT skip any steps. Answer the following questions as you proceed.

1. What is systemic lupus erythematosus (SLE)?

2. From Figure 1 (click on it), what are the four steps of an ELISA protocol?

a. ___________________________________________________________

b. ___________________________________________________________

c. ___________________________________________________________

d. ___________________________________________________________

3. In step 1, you centrifuge the samples. What does a centrifuge do?

4. What are you preparing in step 2? Why are there three different solutions?

5. In steps 3 and 4, you prepare an ELISA plate. What has the ELISA plate been pretreated with? Why?

a. What is the positive control? (Step 4)

b. What is a primary antibody? Please define.

c. What is the negative control? (Step 4)

d. Why is it necessary to have a positive and a negative control? (Step 4)

6. Why incubate the plate in step 5?

7. Next, in step 6, the plate is washed. Why wash the plate?

8. In step 7, a secondary antibody is added. What is a secondary antibody? Please define.

a. What is the attached enzyme in this assay? (Step 7)

b. What is the specific substrate for HRP? What color does it produce? (Step 7)

9. How can the yellow color be quantitatively measured? At what wavelength? (Step 10, in “why”)

10. Record your results. Indicate on this page and on the computer which boxes turned color.

 

A

B

C

+ Control

– Control

 

1:2

 

1:10

 

1:100

11. Did you complete the ELISA correctly? (Yes/No) __________

If yes, proceed to #12 and #14.

If no, proceed to #13 and #14.

12. What do the results indicate about:

Patient A:

Patient B:

Patient C:

13. Explain what you did wrong and what you will need to do next time. For more information, check your printable summary page. Did your incorrect procedure provide you any results? Explain what went wrong.

14. This virtual lab was testing for lupus. How is this same test used to test for the presence of HIV? If the results for an HIV test were the same as in this exercise, what would they indicate about the three patients?

 
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Critical Thinking Discussion

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Students with code numbers starting with a 1 (e.g., 11, 12, 13, etc), must post answers to 2 of the critical thinking questions below. You can only post an answer to a previously answered question if you are correcting an error made by a previous poster.

A person is declared to be dead upon the irreversible cessation of spontaneous body functions brain activity, or blood circulation and respiration. However, only about 1% of a person’s cells have to die in order for all of these things to happen. How can someone be dead when 99% of their cells are still alive?
Explain the difference between a one-celled organism and a single cell of a multicellular organism.
Why would you think twice about ordering from a restaurant menu that lists only the second part of the species name (not the genus) of its offerings? Include an example of why this might be troubling.
Once there was a highly intelligent turkey that had nothing to but reflect on the world’s regularities Morning always started out with teh sky turning light, followed by the master’s footsteps, which were always followed by the appearance of food. Other things varied, but food always followed footsteps. The sequence of events was so predictable that it eventually became the basis fo the turkey’s theory about the goodness of the world. One morning, after more than 100 confirmations fo the goodness of theory, the turkey listened for the master’s footsteps, herd them and had its head chopped off. Any scientific theory is modified or discarded upon discovery of contradictory evidence. The absence of absolute certainty has led some people to conclude that “facts are irrelevant because they can change”. If that is so, should we stop doing scientific research? Why or why not?
In 2005, research Woo-suk Hwang reported that he made immortal stem cells from human patients. His research was hailed as a breakthrough for people affected by degenerative diseases, because stem cells may be used to repair a person’s own damaged cells. Hwang published his results in a peer-reviewed journal. In 2006, the journal retracted his paper after other scientists discovered that Hwang’s group had faked the data. Does this incident show that results of scientific studies cannot be trusted? Or does it confirm the usefulness of a scientific approach, because other scientists discovered and exposed the fraud?

 
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Medical Terms Worksheet

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Orthopedics

Name______________________

1. List alternative therapies for musculoskeletal problems.

2. What is a xyrospasm? What is the Greek derivative of the word?

3. Name and describe the three types of varus conditions.

4. Define the anterior “triangle” of the neck. Give a second example of a body triangle.

5. What are the symptoms of the life-threatening illness caused by the bacteria Clostridium tetani?

6. Ole Worm, a Danish physician (1588-1654) described the wormian bone. Where is this bone found?

7. What is the function of calcium?

8. Divers exposed to the toxin of the stonefish die within days of exposure, a painful, mutilating illness. What toxin does this fish produce and how is it lethal?

9. In radiologic technology, what is the responsibility of the radiographer?

10. Describe the work of a physical therapist.

Orthopedics

Name______________________

1. List alternative therapies for musculoskeletal problems.

2. What is a xyrospasm? What is the Greek derivative of the word?

3. Name and describe the three types of varus conditions.

4. Define the anterior “triangle” of the neck. Give a second example of a body triangle.

5. What are the symptoms of the life-threatening illness caused by the bacteria Clostridium tetani?

6. Ole Worm, a Danish physician (1588-1654) described the wormian bone. Where is this bone found?

7. What is the function of calcium?

8. Divers exposed to the toxin of the stonefish die within days of exposure, a painful, mutilating illness. What toxin does this fish produce and how is it lethal?

9. In radiologic technology, what is the responsibility of the radiographer?

10. Describe the work of a physical therapist.

 
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Lab 5 – Diffusion, Osmosis and Enzymes

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Lab 5 – Diffusion, Osmosis and Enzymes

Objective: The objective of this lab is to introduce you to the concepts of diffusion, osmosis and enzyme activity.

Reading Assignment: Read 4.9 and 5.4; Diffusion and Osmosis and How Enzymes Work from the textbook Essentials of the Living World.

 

For this lab you will need to supply the following materials:

Camera or cell phone (you will upload pictures of your experiment)

Kitchen scale (food scale, it can weigh in either ounces or grams)

Microwave

Liquid measuring cup

Teaspoon

Tablespoon (3 Tsp = 1 Tbs)

Several Drinking cups (1 needs to hold at least 2 cups of liquid)

1 Potato

Knife

Blender or Potato masher

Zip lock sandwich bag (not a freezer bag)

Pancake syrup

Salt

Sugar

Vinegar

Hydrogen Peroxide (H2O2)

 

Solutions to prepare

Sucrose solution – add one tablespoon of sugar to 1 cup of water

 

Exercise 1 – Diffusion

· In this exercise you will observe the diffusion of water through a semipermeable membrane (a plastic bag)

Step 1. Place ÂĽ of a cup of pancake syrup into a plastic zip lock bag and seal the bag.

Step 2. Weigh the bag using a small kitchen scale and record the weight on your worksheet.

Step 3. Fill larger cup (2 cup measuring or equivalent) with 1 cup of water. Record the time.

Step 4. Place the syrup bag in the larger cup and leave overnight (12+ hours).

Step 5. Remove the bag, dry it off and weigh it using a small kitchen scale. Record weight on your worksheet and the time you ended the experiment. (Hint: The bag should gain weight. If it loses weight you may have had a puncture. You must redo this exercise).

Step 6. Take a picture of the completed exercise and upload to the worksheet. Make sure it includes the date, your name and a photo ID.

Exercise 2 – Osmosis

· In this exercise you will determine the result of putting cells into solutions of various tonicity.

Step 1. Cut (2) potato slices: 3 inches long, 1 inch wide, 0.5 inches thick.

Step 2. Prepare two coffee cups with the following solutions:

Cup 1 = add one cup of water

Cup 2 = add one cup of water and 1 teaspoon of salt

Step 3. Add one potato slice to each cup and let stand for 1 hour.

Step 4. Remove potato slices and determine if the slice is crisp or limp. Record your observations of the slices on the data sheet.

Step 5. Take a picture of the completed exercise and upload to the worksheet. Make sure it includes the date, your name, and a photo ID.

Exercise 3 – Enzyme Activity

· In this exercise you will observe factors that affect the enzyme activity of catalase. Catalase is found in potato cells.

H2O2 (hydrogen peroxide) 2 H2O + O2 (oxygen gas) when catalase is present.

Step 1. Place the remaining amount of potato in a blender with two tablespoons of water. Blend to a paste. If you don’t have a blender use a potato masher to make the paste.

Experiment 3-1

Step 2. Transfer one tablespoon of the paste to a small cup and add 1 tablespoon of hydrogen peroxide.

Step 3. Mix together by gently rotating the cup clockwise and record your observations on the data sheet

Step 4. Take a picture of the completed exercise and upload to the worksheet. Make sure it includes the date, your name and photo/student ID.

 

Experiment 3-2

Step 5. Transfer one tablespoon of the paste to a small cup and add 1 tablespoon of sucrose solution.

Step 6. Mix together by gently rotating the cup clockwise and record your observations on the data sheet.

Step 7. Take a picture of the completed exercise and upload to the worksheet. Make sure it includes the date, your name and photo/student ID.

 

Experiment 3-3

Step 8. Transfer one tablespoon of the paste to a small cup and add 1 tablespoon of vinegar (an acid).

Step 9. Next add 1 tablespoon of hydrogen peroxide.

Step 10. Mix together by gently rotating the cup clockwise and record your observations on the data sheet.

Step 11. Take a picture of the completed exercise and upload to the worksheet. Make sure it includes the date, your name and photo/student ID.

 

Experiment 3-4

Step 12. Transfer one tablespoon of the paste to a small cup and add 1 tablespoon of water.

Step 13. Microwave on high for 1 minute.

Step 14. Remove from microwave (careful it’s hot) and allow 5 minutes to cool.

Step 15. Next add 1 tablespoon of hydrogen peroxide.

Step 16. Mix together by gently rotating the cup clockwise and record your observations on the data sheet.

Step 17. Take a picture of the completed exercise and upload to the worksheet. Make sure it includes the date, your name and photo/student ID.

 
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