Dual dean entrainment with volume ratio modulation for efficient droplet co-encapsulation: extreme single-cell indexing

The future of single cell diversity screens involves ever-larger sample sizes, dictating the need for higher throughput methods with low analytical noise to accurately describe the nature of the cellular system. Current approaches are limited by the Poisson statistic, requiring dilute cell suspensions and associated losses in throughput. In this contribution, we apply Dean entrainment to both cell and bead inputs, defining different volume packets to effect efficient co-encapsulation. Volume ratio scaling was explored to identify optimal conditions. This enabled the co-encapsulation of single cells with reporter beads at rates of similar to 1 million cells per hour, while increasing assay signal-to-noise with cell multiplet rates of similar to 2.5% and capturing similar to 70% of cells. The method, called Pirouette coupling, extends our capacity to investigate biological systems.

Automated summary

The future of single cell diversity screens requires larger sample sizes and higher throughput methods to accurately describe cellular systems. The author utilized Dean entrainment to efficiently co-encapsulate single cells with reporter beads at high rates, improving assay signal-to-noise and capturing a high percentage of cells.