Real-Time Deformability Cytometry: High-Throughput Mechanical Phenotyping for Changes in Cell Function

Abstract

Changes in cell function are often accompanied by alterations of the intrinsic cell structure particularly the cytoskeleton. This leads to distinct mechanical changes. For example, cells become softer during malignant transformation and stiffer during differentiation. Exploiting the mechanical phenotype of cells as an inherent, label-free marker requires a high-throughput and robust measurement technique. Here, we introduce real-time deformability cytometry (RT-DC) for mechanical single cell classification of heterogeneous cell populations at rates of several hundred cells per second in real-time. Performing RT-DC on primary human hematopoietic stem cells and mature blood cells we demonstrate its capability to detect lineage and source specific mechanical phenotypes. We also find that different stages of the cell cycle possess a unique mechanical fingerprint allowing the distinction between cells in G2 and M phase. In summary, RT-DC represents a novel flow cytometric approach that enables the translation of mechanical phenotyping from basic research into applications in biology and medicine.