Tilting against windmills: Clarity in spectral panel design without losing the battle for biological resolution

Sponsored by: Thermo Fisher Scientific

C.E. Credits: P.A.C.E. CE ｜ Florida CE

Speaker

Event Date & Time

Date: October 12, 2022, October 13, 2022

Time: 8:30pm (PDT), 11:30pm (EDT), 5:30am (CEST), 11:30am (SGT)

Abstract

High dimensional full spectrum flow cytometry holds the promise of addressing long-held questions in cellular biology. Fluorescent dye performance heavily impacts both the quality and the dimensionality of flow cytometry experiments. The development of fluorescent labels engineered with targeted excitation and emission spectra using the Invitrogen™ Phiton™ DNA platform has allowed the generation of flow cytometry panels with the goal of reducing spectral spread and increasing biological resolution. In this tutorial, we will take a deep dive into the data generated using a high parameter panel while leaving available placeholders for commonly used fluorophores. We will also discuss the iterative panel design process and identify problematic fluorophore combinations as well as best practices for panel design practices that enable greater biological resolution.

Learning Objectives:

Discuss similarities and differences in conventional and spectral flow cytometry panel design
Construct spectral flow cytometry panels to avoid cross-excitation and spectral spread
Utilize similarity index and complexity score to select compatible dyes for use in spectral flow cytometric panels
Evaluate existing flow cytometry panels using the spectral spread matrix (SSM) to identify problematic dyes and improve biological resolution

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