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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 PHYSICS


Included in this lesson plan:

  • This overview document

  • A PowerPoint presentation with lectures for each day

  • A document outlining student activities

  • Links to resources

  • The beginning and advanced FlowJo tutorials (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:

  • Explain what a Flow Cytometer is used for

  • Identify and explain the basic components of a Flow Cytometer

  • Explain what gating is

  • Do basic analysis in FlowJo


By the end of 2 week lesson, they should learn:

  • How the fluidic, optics, electronics, optical measurement, and data analysis systems of a flow cytometer work

  • The applications of flow cytometry


Students should already know (or may need a very brief overview):

  • What a cell is

  • What T-Cells are in general terms

  • What an electron is

  • Basic principles of graphing (what a histogram is, how to read graphs based on axes, etc)


Some Physics knowledge that can be referenced during this lesson or that can be used as accompanying lessons:

  • Optics (lasers, wavelength, frequency, internal reflection, defraction, color, fiber optics, lenses, photons)

  • Hydrodynamics (pressure)

  • Electronics (voltage, charge, cathodes, electrons)


An Overview of the PowerPoint lectures and activities:

  • Ask who in the class knows someone with cancer, HIV, or AIDS

  • What if we want to identify diseased cells? How would we distinguish between different types of cells?

  • The answer: cytometry!

  • What is Cytometry? Let’s break the word down:

    • Cyto = Cell

    • Metry = Measure

    • Therefore; literally: Cell-measure

    • What it measures (for instance, not solid tissues): size, granularity, internal complexity

  • 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)

  • Briefly, how the body recognizes cells (immune system, T cells, etc)

  • A modern procedure: Flow Cytometry

  • What we use Flow Cytometry for

  • Step-by-step through flow cytometry

    • The three parts: Fluidics, Optics, Electronics

    • Add a compound that marks the cells (compare to: dyes, stains, reagents) (maybe look at structure of the fluorescence molecules)

    • Get cells into a single-file line

    • Shine a laser at them that excites the marker

  • Now explain these parts in more detail. Their functions, etc.

    • The Fluidics using the cell stream and pressures of hydrodynamic focusing of the sheath fluid

    • The Optics with lasers and sensors, excitation and flourescence

    • Electronics

    • Analysis, with compensation, gating, etc

  • Record the reflected light and its frequency makeup

  • 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.

  • 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?)


Follow with the PowerPoint presentation and make sure to look at the notes included at the bottom of slides


If you finish early on the daily lectures, you can look at some of the links below, especially the BDBioscience link.


ACTIVITIES: (see worksheet)


T-Shirt Flow Cytometry overview

Candy sorting to show gating


Resources:

(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

 

 

Contents are solely the responsibility of the authors and do not necessarily represent the official views of the NIH.
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