CYTO 2026 Scientific Tutorial: Approaches to Spectral Unmixing Challenges, Journey of a Detective

Approaches to Spectral Unmixing Challenges, Journey of a Detective

Presenters:
Sara Garcia-Garcia, MSc, Technical Director, Microscopy and Flow Cytometry Facility, Amsterdam UMC
Laura Ferrer Font, PhD, Scientific Solutions Manager, BD Biosciences

Abstract:

Do you have what it takes to become a spectral detective? Are you tired of those unresolvable unmixing mysteries that keep haunting your plots? Grab your trench coat and magnifying glass, because two seasoned investigators are on the case, and they want you on the team. Each case hides clues about spectral unmixing challenges, and your mission is to uncover the truth.

Which will be your role? Analyze evidence, apply your unmixing skills, and solve the cases step by step. By the end, you’ll have the tools and confidence to tackle even the most stubborn spectral mysteries.

With a mix of hard-boiled humor and cytometric wisdom, we’ll expose the good, the bad, and the ugly of spectral unmixing. Because in spectral cytometry, every photon leaves a trace, and every bad design leaves a body (of messy data).  Join us on an investigative adventure where science meets mystery!

Learning Objectives:

Recognize common pitfalls in spectral panel design and why some fluorochrome choices can never be 'resolved'.
Understand how single-stain control errors (e.g., spectral signatures mismatch or AF mismatches) impact unmixing quality.
Learn strategies for dealing with autofluorescence.
Develop practical troubleshooting skills to spot, diagnose, and solve spectral unmixing issues.
Recognise best practices to make unmixing reproducible across experiments.

Keywords: Fundamental Cytometry Concepts, Experimental Design & Controls, Unmixing, Spectral Flow Cytometry

CMLE Credit: 1.5