Rockets Into The Unknown:
Diagnosing Multiple Sclerosis

Developed by Regina Lindsey-Lynch, Larry Osmundson, Bruce Price,
and Martha Thompson
© American Association of Immunologists 1992

Focus
This laboratory exercise is designed to show a medical application of basic immunology. The technique of rocket electrophoresis visually illustrates the antigen-antibody precipitation reaction. It can be used to measure the level of antibody (IgG) in body fluids, thereby aiding in diagnosis of certain diseases such as multiple sclerosis (MS).

Description
Laboratory exercise 1 is an exploratory activity that introduces the technique of electrophoresis. It is designed to allow students to become familiar with both the equipment and the concept of how substances move through an agarose gel. In this activity dyes are electrophoresed. Each dye can be seen as a different color on the agarose gel.

Laboratory exercise 2 uses the technique of rocket electrophoresis to measure the "simulated IgG" in "simulated cerebral spinal fluid (CSF)." This technique is performed in an agarose gel containing an antibody that will react with the "IgG." When the "CSF" is applied to the gel, a rocket-shaped precipitate is formed. The height of the rocket is approximately proportional to the concentration of antigen. Elevated levels of IgG (the antigen) indicate that MS is a possible diagnosis.

Laboratory exercise 2A is a qualitative investigation with one control and two unknowns. Exercise 2B involves the construction of a standard curve and is quantitative in its approach.

Objectives
Upon completion of these lessons, students should be able to:

  1. Define these terms and use them in context: multiple sclerosis, autoimmune, remission, antigen, antibody, electrophoresis.
  2. Discuss the nature of the antigen-antibody complex as it applies to the formation of the rocket.
  3. Explain the medical application of the immunological technique of rocket electrophoresis.
  4. Display competence in the laboratory techniques for micropipetting and gel electrophoresis.
  5. Describe the process and purpose of gel electrophoresis.
  6. Compare unknown samples with a standard curve (Exercise 2B only).
  7. Draw conclusions based on analysis of experimental results.

Suggested Sequence
Instructional time: Five 45-50 minute class periods

Day One: Introduction Activities

  1. Teacher will demonstrate the proper and safe use of the materials and equipment.
  2. Students will practice using micropipettors.
  3. Students will prepare slides for the experiment.
  4. Teacher will discuss separation by electrophoresis.

Day Two: Exploratory Activities

  1. Students will perform Experiment 1.
  2. Students will observe, record data, and draw conclusions.
  3. Teacher will demonstrate precipitation reaction and discuss the formation of the antigen-antibody complex and rocket.
  4. Class will analyze and discuss data.

Day Three: Introduction to MS and Preparation for Experiment 2

  1. Class will discuss MS.
  2. Students will watch video on MS.
  3. Students will prepare slides and materials for Experiment 2.

Day Four:. Experiment 2: Rocket Electrophoresis

  1. Students will perform the electrophoresis.
  2. Class will discuss data interpretation and application.

Day Five: Data Analysis Activities

  1. Students will develop slides from Experiment 2.
  2. Class will analyze and discuss data.