High-Dimensional Cytometry in the Spectral Era: New Metrics to Design a 50-Color Panel

About the Speakers

Florian Mair - Scientific Director of Flow Cytometry Core Facility, ETH Zurich, Switzerland

Florian is currently working as the Scientific Director of the Flow Cytometry Core Facility and Senior Scientist Immunology at ETH Zurich, Switzerland. After graduating with an MSc in Molecular Biology from the University of Vienna, he received his PhD in Molecular Life Sciences from the University of Zurich in 2014, and spent several years at the Fred Hutchinson Cancer Center in Seattle, USA. During the past decade, he has been involved extensively with different high-dimensional cytometry platforms as well as scRNA-sequencing techniques, including multi-omic methods. His main scientific interests are the function of dendritic cells and T cells in non-lymphoid tissues during health and disease, teaching best practices in cytometry and developing new flow cytometry panels and approaches.

Webinar Summary

To understand the function of the human immune system it is imperative to capture as much information as possible from often size-limited human samples. To do so, we have developed the first 50-color spectral flow cytometry panel to comprehensively study the functional state of the human immune system in PBMCs and tissue samples. The panel contains lineage markers for all major immune cell subsets, and an extensive set of phenotyping markers focused on the activation and differentiation status of the T cell and dendritic cell (DC) compartment. The panel has also been tested to be suitable for cell sorting, which allows very fine-grained isolation of immune subsets utilizing 50 markers at the same time. 

To establish such a complex panel, we utilized a new metric termed unmixing spreading error, that evaluates the fluorochrome-specific increase in background that is inherently generated when unmixing highly complex panels. In this presentation we show the systematic workflow we used to develop the panel, how we evaluated panel performance, and how OMIP-102 was developed to its final state.

Learning Objectives

Panel Design, Spreading Error, Unmixing, OMIP design

Who Should Attend

Everyone interested in high-dimensional flow cytometry panels and OMIP-102n

CMLE Credit: 1.0