Large Particle Fluorescence-Activated Cell Sorting Enables High-Quality Single-Cell RNA Sequencing and Functional Analysis of Adult Cardiomyocytes

T he advent of single-cell RNA sequencing (scRNA-seq) has offered great opportunities for studying cardiac pathology at single-cell resolution. To date, however, scRNA-seq of adult cardiomyocytes remains limited owing to technical difficulties in single cardiomyocyte isolation. Adult cardiomyocytes are large (125x25 mu m), rodshaped cells whose structure precludes isolation by fluorescence-activated cell sorting (FACS) or commercial single-cell microfluidic platforms. 1 Here, we report on successful isolation of viable single cardiomyocyte through use of large particle FACS (LPFACS), designed for the isolation of large objects, including worm/fly embryos and cell clusters. Our study demonstrates that LP-FACS enables rapid, high-throughput isolation of cardiomyocytes. To generate viable cardiomyocytes,