Tech exchange with leica: overcoming the high multiplexing barrier: 3d spatial omics in one go, powered by leica microsystems"

The STELLARIS confocal platform offers significant advancements for high-multiplex strategies, integrating a tunable white light laser (WLL) with an excitation range from 440 nm to 790 nm and up to five highly sensitive, photon-counting spectral detectors. This configuration enables complete flexibility in detection from 410 nm to the near-infrared (NIR) spectrum (1), accommodating a broad array of fluorophores and optimizing their combinations for complex imaging tasks.

In this presentation, we will introduce the new SpectraPlex 3D high-multiplex solution for the STELLARIS confocal platform (2, 3). We will detail the experimental design and considerations for executing high-multiplex experiments with 15+ fluorophores on a single sample, including the selection of optimized panels and the inherent challenges associated with these approaches. We will address the technical aspects of imaging high-multiplex samples in a single round of labeling and imaging, demonstrating its application for 3D spatial omics. 

Our aim is to provide a comprehensive understanding of how these technologies can be leveraged to generate detailed spatial omics data, thereby advancing the field of spatial biology. 

1. The Power HyD family of detectors. Schweikhard et al., Nature Methods (2022). DOI: d42473-020-00398-0.
2. 3D multiplexing imaging in cancer immunology. Kunz L., Speziale D., et al., Nat. Methods (2024). DOI: d42473-024-00260-7
3. SpectraPlex: A powerful toolbox for advanced 3D high-multiplex imaging. Roberti, M. J., Hecht F. et al., Nat. Methods in press (2024). DOI: d42473-024-00262-5