How to overcome the high multiplexing barrier

17:10 – 18:10 BST, 5 June 2024 ‐ 1 hour

Leica Microsystems

Luis Alvarez1, Irmtraud Steinmetz1, Tatjana Straka1, Susanne Holzmeister1, Quyen Tran2, Julia Roberti1

1Leica Microsystems, Germany. 2Leica Microsystems, USA

Workshop Room 11C

The STELLARIS confocal platform is particularly well suited for high multiplexing strategies as it combines a freely tunable white light laser (WLL) (440nm - 790nm) excitation with up to 5 highly sensitive spectral detectors for complete flexibility of detection from 410 nm up to the NIR range1. This unique combination allows it to fit a large palette of fluorophores and to optimise any possible combination.

In this workshop, we will elaborate on the experimental design and considerations needed to perform high multiplexing imaging experiments beyond 10 fluorophores on a single sample2. We will discuss appropriate fluorophore panels, details of sample preparation and the challenges and tools required for imaging. 

Gathering insights from these rich and detailed images requires advanced image analysis. We will also showcase how Aivia’s advanced tools, many powered by AI, can support this requirement.

References 

  1. Application Note: The Power HyD family of detectors, Schweikhard et al. (2022). Nature Methods, doi: d42473-020-00398-0 
  2. Multiplexing through Spectral Separation of 11 Colors, Hoffmann S. et al. (2022). https://www.leica-microsystems.com/science-lab/life-science/multiplexing-through-spectral-separation-of-11-colors/