High-Dimensional Cell Sorting Utilizing Real-Time Spectral Unmixing With The Bigfoot Spectral Cell Sorter

By Alan Saluk, Brian Seegers, Brian Monteverde, and Nicholas Rohrbacker

The rapid advance of flow cytometry over the past two decades has been an incredible asset to the cell research community. The convergent evolution of digital electronic acquisition, solid-state laser excitation, refined emissions collection optics, advanced hardware automation, and fluorophore development has increased capabilities from 4 to 40 colors, allowing more data from each sample. Analytical flow cytometry and multicolor phenotyping panels have been helpful in establishing the presence of cell subsets. However, segregation of specific cell subsets for molecular profiling and sequencing has been problematic because cell sorters have lacked the technical sophistication to isolate cell types found in complex multicolor spectral panels and sort them in real time.

The Invitrogen™ Bigfoot Spectral Cell Sorter was designed to isolate cell types found in complex multicolor spectral panels and sort them in real time by pairing the power of full-spectrum flow cytometry with real-time sorting to extend the frontiers of cell type discovery. We demonstrate the capability of the Bigfoot Spectral Cell Sorter using a 23-color panel designed for isolating Siglec-expressing cell subpopulations, which are expressed on most white blood cells of the immune system and play critical roles in immune cell signaling.