Panel design – A Practical Guide for Successful Fluorescent Cytometry Panels from 10-40 Parameters
Immunologist | Cytometry Specialist
Fred Hutchinson Cancer Research Center
About the Presenter
Florian graduated with a PhD from the University of Zurich, Switzerland, in 2014 and is currently working at the Fred Hutchinson Cancer Research Center in Seattle, USA, as an immunologist. During the past decade, he has been involved extensively with different cytometry platforms (conventional, spectral and mass cytometry) as well as scRNA-seq techniques and developed an interest in applying novel analysis approaches for single cell data. He has been actively engaged in teaching flow cytometry courses, including systematic panel design and analysis of high-dimensional cytometry experiments. Florian is currently an ISAC Marylou scholar.
Over the past decade, technical improvements and new reagents have permitted fluorescent-based flow cytometry assays to measure up to 40 parameters. These complex assays require robust controls and thorough experimental planning, but there are currently few resources that provide a systematic approach for reliable panel design. Also, historical notions as to how fluorophores and controls should be chosen are sometimes at odds with the reality of modern panel design.
In this webinar, we will provide a practical guide for successful fluorescent panel design for any complex panel from 10-40 (or more) parameters, both for conventional compensation-based as well as spectral cytometry.
Specifically, we will cover the following topics:
- Brief overview of signal detection in conventional and spectral flow cytometers
- The concept and underlying cause of spreading error (SE)
- How the spillover spreading matrix (SSM) can be efficiently used to guide panel design
- Relevant relationships between SE, fluorophore brightness and antigen expression level
- Step-by-step approaches towards building a new panel
- An overview of essential controls and typical caveats
This webinar is back-to-back with a webinar by Thomas Liechti (NIH, USA), who will be talking about how to best design and use complex panels for large study cohorts (100s-1000s of samples), including the use of appropriate controls and analysis approaches.
After attending these two back-to-back webinars, participants should have gained a solid base for tackling high-dimensional immunophenotyping assays on a variety of platforms, thus minimizing trial-and-error experiences and wasted experiments. Also the attendees will learn specific strategies for generating reproducible and high-quality data in large patient cohorts.
Who Should Attend
Anyone with an interest in efficient panel design: Immunologists, Scientists of any field doing polychromatic flowcytometry, SRL-users and SRL-leaders.