Biochemistry

Welcome to the Tree Star-sponsored flow cytometry lesson plans for high school students and educators. This curriculum was put together with funds from Grant #024094 from NIAID.

Students, just click on your subject below to download your lesson plan and resources.

Educators will find it helpful to read the Educational Resources overview.

Flow Cytometry for High Schools

With a focus on BIOCHEMISTRY/IMMUNOLOGY

Included in this lesson plan:

PowerPoint
Worksheets for T-Shirt activity and candy sorting activity
Tutorial for in-class FlowJo project
You can go online to http://www.flowjo.com/ to download the beginning and advanced tutorials. You can either use these as teacher resources or have students complete them.

Objectives:

By the end of this two-week lesson, students should be able to:

Understand how the body’s immune system recognizes cells
Understand basic flow cytometry (including parts of a cytometer)
Understand how FlowJo can be used to diagnose HIV/AIDS
Be able to analyze data in FlowJo and come to a conclusion

Basic Overview with PowerPoint

Ask who in the class knows someone with cancer, HIV, or AIDS. How do we diagnose these diseases?
What if we want to identify diseased cells? How would we distinguish between different types of cells?
The answer: flow cytometry!
What is Flow Cytometry? Let’s break the word down:
- Flow = fluid movement
- Cyto = Cell
- Metry = Measure
- Therefore; literally: Cell measure through fluid movement
- What it measures (for instance, not solid tissues): size, granularity, internal complexity, many other things
How do we “measure” cells? How do we normally gather information about things we see around us every day?
- The human senses
  - Sight (Light)
  - Sound (Vibrations)
  - Touch (Texture)
  - Smell
  - Taste (molecule makeup)

How the body recognizes cells:

Body has antibodies that recognize a certain type of cell.
Antigens (what an antibody attaches to) have certain specific receptors (link to lock and key/proteins).
Antibodies recognize these receptors (called epitopes) and bind to them.
Flow cytometry uses fluorescent antibodies to distinguish between cells.

Step-by-step through flow cytometry

Brief overview of the three parts: Fluidics, Optics, Electronics
- (insert diagram here)
Add a fluorescent antibody (compare to: dyes, stains, reagents) (maybe look at structure of the fluorescence molecules)
Get cells into a single-file line (in fluid)
Shine a laser at them one at a time
Antibody fluoresces (emits certain colors of light)
Sensors record the reflected light and its frequency makeup, and send the data to a computer to be analyzed

Flowjo Activity, See worksheet

Certain markers bind with certain surface proteins
By looking at the light, we can tell which markers bound to the cells, and by extension, what kind of cells they are.
Connect flow cytometry to diagnosing disease; how do we do it? (Note: this is where you would connect to a cell unit, integrating previous knowledge such as surface proteins and the lock-and-key model)
- Many diseases have specific epitopes on the cellular level; to find out if someone has that disease, you look for the fluorescent peptide (antibody) that binds to that epitope,
Now we start the activity using the actual Flowjo software.

General introduction to the software for the activity (give basic and advanced tutorial to teachers; they can do what they want with it)
Introduction of activity and goals
- Find out who has AIDS
Do the activity
Options for conclusion of project:
- Write a letter to the patient/’s family explaining to a lay person the results of the test
- Write a lab report explaining the procedure and how you arrived at your conclusion
- Do further analysis on the data and create a report on your findings
- Research another aspect of flow cytometry and give a presentation with visual aids (extra credit?)

Possible projects/activity:

See worksheets for the T-Shirt and candy sorting activities

Immune Response Teams: Students are broken up into collaborative groups. Each student in the group is responsible for one cell type. After completing this activity, students teach their peers about their cell. In groups, or individually, students create story boards, cartoons or books that show their understanding of cell players in the immune system.

Resources (you may wish to go over some of these in class with the students):

(for an overview video included in the Powerpoint) http://www.youtube.com/watch?v=nAfL4FXju1s

(an in-depth intro) http://www.abdserotec.com/uploads/Flow-Cytometry.pdf

(very helpful overview of the different componants) http://www.bdbiosciences.com/support/training/itf_launch.jsp

(a more basic, easy to understand introduction) http://www.scq.ubc.ca/flow-cytometry-a-technology-to-count-and-sort-cells/

(A helpful, very visual guide) http://www.unsolvedmysteries.oregonstate.edu/flow_04

http://www.unsolvedmysteries.oregonstate.edu/flow_06