A superior dead cell removal platform using Akadeum’s BACS microbubbles

Abstract

Abstract Single cell sequencing (SCS) is one of the most commonly used analytical techniques for cellular characterization and has become a mainstay in immune profiling. As SCS has gained in popularity and improved in sensitivity, it has become apparent that ambient nucleic acids can have a major impact on the quality and reproducibility of obtained SCS results. Since the bulk of ambient nucleic acids in a sample come from dead or dying cells that have lost membrane integrity, one way to dramatically reduce ambient nucleic acids and decrease background noise in SCS is to remove dead cells from the sample prior to sequencing. To address the challenge presented by contaminating dead cells, Akadeum has developed a Buoyancy Activated Cell Sorting (BACS™) microbubble approach for depleting dead cells from biological samples. Leveraging the power of gravity to drive self-isolation, Akadeum’s BACS microbubbles represent some of the gentlest isolations available; therefore, they are ideally suited for applications where cell viability is integral to success of the experiment. Additionally, BACS microbubbles from Akadeum are quick and easy to use since they require only minimal manipulation and do not rely on any extra equipment or consumables. Depletion of dead cells is achieved through the selective capture of cells with exposed phosphatidylserine with Annexin V conjugated BACS microbubbles. Once mixed with the sample, the Annexin V BACS microbubbles capture dead cells and float them to the sample surface for removal leaving behind healthy and untouched cells. Samples exposed to the Annexin V BACS microbubbles demonstrated significantly reduced dead cell populations and ultimately yielded more reliable results.