Fluorescent Proteins in Flow Cytometry

The Presenter

Teresa Hawley
Flow Cytometry Core Facility
George Washington University

Bill Telford
NCI Flow Cytometry Core Laboratory
National Institute of Health

John Nolan, PhD
Professor
Scintillon Institute

Session Summary

In a scientific tutorial in 2011, Bill Telford covered the fluorescent proteins (FPs) available at the time. As a follow up, we will focus on recent developments in the field, including new FPs, new lasers, and the advances made in detecting FPs using spectral flow cytometry. We will discuss considerations for practical use of FPs and make recommendations for choice of FPs in flow cytometry. We will analyze FP data using fluorescence compensation for conventional flow cytometry and unmixing algorithms for spectral flow cytometry. Attendees are encouraged to bring along their own data. Upon completion of this tutorial, the attendee will have an overview of the properties of commonly used FPs, the pertinent information to create optimal combinations of FPs, the knowledge to devise feasible detection strategies, and the skill to analyze FP data.

Session Outline

1. Basics/Overview.
2. Timeline of major developments in FPs.
3. List of commonly used FPs.
4. Considerations for practical use of FPs.
5. Recommendations for choice of FPs in flow cytometry.
6. Strategies for simultaneous detection of multiple FPs.
7. Matching lasers with FPs.
8. Conventional vs spectral flow cytometry of FPs.
9. Flow cytometer calibration using FP beads.
10. Fluorescence compensation in conventional flow cytometry.
11. Unmixing algorithms in spectral flow cytometry.
12. Display of properly compensated FP data.
13. Discussion.

CMLE Credit: 1.0